Difference: EukExpFaqs (1 vs. 27)

Revision 2711 Dec 2009 - Main.DavidCowburn

 
Q A
What happened to the proposal? As of 21 May, it is accepted at NIH and assigned to a special study section. The initial peer review should be completed by the end of November 2009 and a funding decision made shortly after the appropriate National Advisory Group meets in January 2010.
We are assuming that we could not do yeast or Dictyostelium expression - is this correct? It's not clear that the sophisticated procedures available would significantly enhance throughput for yeast. This is something to be explored when the system is established. For Dicty, the issue would presumably be that the current setup is for free cells in suspension, not surface-associated, and that may not be what's needed for your Dicty systems. Finally, there would have to be an operational view of how many different cell systems could be routinely incorporated without cross contamination issues.
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 μ g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioreactors/flasks is a question of cost effectiveness, that users could self determine.
how would this facility work- we would be able to provide bacculoviral constructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The initital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this.
Over the long term, it will likely operate on a fee-for-service basis.
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm
I do NMR and I dont think this could help me isotopically label. Any comments? 1) finding the right construct in a eukaryotic screen is likely to be generally useful and be a lead for cell free or prokaryotic expression , and can identify useful complexes, post-translation mods etc.
2) for a few, high profile, cases, the economics of euk expression may be manageable once you know you have high expression. Refs -- LIBRARY:OhkiDohi08.pdf etc . But realistically we'd have to say this is a research objective once the system is in place. Note that we will have the 1.7 mm TCI 800 probe so small quantities of labeled material are useful.
3) You can think big, e.g. inducible expression of unlabeled materials interacting with injected/ transported labeled materials a la recent Nature papers.
I have another question Please add it to the comment box below
Changed:
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Revision 2605 Oct 2009 - Main.DavidCowburn

 
Q A
What happened to the proposal? As of 21 May, it is accepted at NIH and assigned to a special study section. The initial peer review should be completed by the end of November 2009 and a funding decision made shortly after the appropriate National Advisory Group meets in January 2010.
We are assuming that we could not do yeast or Dictyostelium expression - is this correct? It's not clear that the sophisticated procedures available would significantly enhance throughput for yeast. This is something to be explored when the system is established. For Dicty, the issue would presumably be that the current setup is for free cells in suspension, not surface-associated, and that may not be what's needed for your Dicty systems. Finally, there would have to be an operational view of how many different cell systems could be routinely incorporated without cross contamination issues.
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 μ g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioreactors/flasks is a question of cost effectiveness, that users could self determine.
how would this facility work- we would be able to provide bacculoviral constructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The initital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
Changed:
<
<
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
Over the long term, it will likely operate on a fee-for-service basis.
>
>
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this.
Over the long term, it will likely operate on a fee-for-service basis.
 
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm
Changed:
<
<
I do NMR and I dont think this could help me isotopically label. Any comments? 1) finding the right construct in a eukaryotic screen is likely to be generally useful and be a lead for cell free or prokaryotic expression , and can identify useful complexes, post-translation mods etc.
2) for a few, high profile, cases, the economics of euk expression may be manageable once you know you have high expression. Refs -- LIBRARY:OhkiDohi09.pdf etc . But realistically we'd have to say this is a research objective once the system is in place. Note that we will have the 1.7 mm TCI 800 probe so small quantities of labeled material are useful.
3) You can think big, e.g. inducible expression of unlabeled materials interacting with injected/ transported labeled materials a la recent Nature papers.
>
>
I do NMR and I dont think this could help me isotopically label. Any comments? 1) finding the right construct in a eukaryotic screen is likely to be generally useful and be a lead for cell free or prokaryotic expression , and can identify useful complexes, post-translation mods etc.
2) for a few, high profile, cases, the economics of euk expression may be manageable once you know you have high expression. Refs -- LIBRARY:OhkiDohi08.pdf etc . But realistically we'd have to say this is a research objective once the system is in place. Note that we will have the 1.7 mm TCI 800 probe so small quantities of labeled material are useful.
3) You can think big, e.g. inducible expression of unlabeled materials interacting with injected/ transported labeled materials a la recent Nature papers.
 
I have another question Please add it to the comment box below

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Revision 2521 May 2009 - Main.DavidCowburn

 
Q A
Added:
>
>
What happened to the proposal? As of 21 May, it is accepted at NIH and assigned to a special study section. The initial peer review should be completed by the end of November 2009 and a funding decision made shortly after the appropriate National Advisory Group meets in January 2010.
 
We are assuming that we could not do yeast or Dictyostelium expression - is this correct? It's not clear that the sophisticated procedures available would significantly enhance throughput for yeast. This is something to be explored when the system is established. For Dicty, the issue would presumably be that the current setup is for free cells in suspension, not surface-associated, and that may not be what's needed for your Dicty systems. Finally, there would have to be an operational view of how many different cell systems could be routinely incorporated without cross contamination issues.
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 μ g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioreactors/flasks is a question of cost effectiveness, that users could self determine.
Deleted:
<
<
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
 
how would this facility work- we would be able to provide bacculoviral constructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The initital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
Changed:
<
<
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
>
>
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
Over the long term, it will likely operate on a fee-for-service basis.
 
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm
Deleted:
<
<
I think it might be a stretch for me to qualify as a major user, but I can certainly put together a description as an "other user." What's the cross over point? Assuming that cost isn’t a factor (see above) if you can think of 1000 constructs / year to test in your current and hopefully expanded programs going forward -- that’s about the 5% line for a major user. For prokaryotic expression, it’s not unusual for NMR people to make 20-30 constructs per structural target. And this cycle is largely limited by how long it takes to do the structures. For functional studies (especially in any high throughput mode), 50 targets a year sounds very modest implying about 1000 constructs
How did you make up the numbers in the q above? GNF claim 600 constructs/ week. We will cautiously plan for 400 and 50 weeks / y output == 20,000 /y . So a 5% major user is 1,000/y
 
I do NMR and I dont think this could help me isotopically label. Any comments? 1) finding the right construct in a eukaryotic screen is likely to be generally useful and be a lead for cell free or prokaryotic expression , and can identify useful complexes, post-translation mods etc.
2) for a few, high profile, cases, the economics of euk expression may be manageable once you know you have high expression. Refs -- LIBRARY:OhkiDohi09.pdf etc . But realistically we'd have to say this is a research objective once the system is in place. Note that we will have the 1.7 mm TCI 800 probe so small quantities of labeled material are useful.
3) You can think big, e.g. inducible expression of unlabeled materials interacting with injected/ transported labeled materials a la recent Nature papers.
I have another question Please add it to the comment box below

Back to: GetAllFAQs

Revision 2429 Apr 2009 - Main.DavidCowburn

 
Q A
We are assuming that we could not do yeast or Dictyostelium expression - is this correct? It's not clear that the sophisticated procedures available would significantly enhance throughput for yeast. This is something to be explored when the system is established. For Dicty, the issue would presumably be that the current setup is for free cells in suspension, not surface-associated, and that may not be what's needed for your Dicty systems. Finally, there would have to be an operational view of how many different cell systems could be routinely incorporated without cross contamination issues.
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 μ g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioreactors/flasks is a question of cost effectiveness, that users could self determine.
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
how would this facility work- we would be able to provide bacculoviral constructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The initital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm
I think it might be a stretch for me to qualify as a major user, but I can certainly put together a description as an "other user." What's the cross over point? Assuming that cost isn’t a factor (see above) if you can think of 1000 constructs / year to test in your current and hopefully expanded programs going forward -- that’s about the 5% line for a major user. For prokaryotic expression, it’s not unusual for NMR people to make 20-30 constructs per structural target. And this cycle is largely limited by how long it takes to do the structures. For functional studies (especially in any high throughput mode), 50 targets a year sounds very modest implying about 1000 constructs
How did you make up the numbers in the q above? GNF claim 600 constructs/ week. We will cautiously plan for 400 and 50 weeks / y output == 20,000 /y . So a 5% major user is 1,000/y
Changed:
<
<
I do NMR and I dont think this could help me isotopically label. Any comments? 1) finding the right construct in a eukaryotic screen is likely to be generally useful and be a lead for cell free or prokaryotic expression , and can identify useful complexes, post-translation mods etc.
2) for a few, high profile, cases, the economics of euk expression may be manageable once you know you have high expression. Refs -- LIBRARY:OkhiDohi09.pdf etc . But realistically we'd have to say this is a research objective once the system is in place. Note that we will have the 1.7 mm TCI 800 probe so small quantities of labeled material are useful.
3) You can think big, e.g. inducible expression of unlabeled materials interacting with injected/ transported labeled materials a la recent Nature papers.
>
>
I do NMR and I dont think this could help me isotopically label. Any comments? 1) finding the right construct in a eukaryotic screen is likely to be generally useful and be a lead for cell free or prokaryotic expression , and can identify useful complexes, post-translation mods etc.
2) for a few, high profile, cases, the economics of euk expression may be manageable once you know you have high expression. Refs -- LIBRARY:OhkiDohi09.pdf etc . But realistically we'd have to say this is a research objective once the system is in place. Note that we will have the 1.7 mm TCI 800 probe so small quantities of labeled material are useful.
3) You can think big, e.g. inducible expression of unlabeled materials interacting with injected/ transported labeled materials a la recent Nature papers.
 
I have another question Please add it to the comment box below

Back to: GetAllFAQs

Revision 2329 Apr 2009 - Main.DavidCowburn

 
Q A
We are assuming that we could not do yeast or Dictyostelium expression - is this correct? It's not clear that the sophisticated procedures available would significantly enhance throughput for yeast. This is something to be explored when the system is established. For Dicty, the issue would presumably be that the current setup is for free cells in suspension, not surface-associated, and that may not be what's needed for your Dicty systems. Finally, there would have to be an operational view of how many different cell systems could be routinely incorporated without cross contamination issues.
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 μ g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioreactors/flasks is a question of cost effectiveness, that users could self determine.
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
how would this facility work- we would be able to provide bacculoviral constructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The initital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm
I think it might be a stretch for me to qualify as a major user, but I can certainly put together a description as an "other user." What's the cross over point? Assuming that cost isn’t a factor (see above) if you can think of 1000 constructs / year to test in your current and hopefully expanded programs going forward -- that’s about the 5% line for a major user. For prokaryotic expression, it’s not unusual for NMR people to make 20-30 constructs per structural target. And this cycle is largely limited by how long it takes to do the structures. For functional studies (especially in any high throughput mode), 50 targets a year sounds very modest implying about 1000 constructs
How did you make up the numbers in the q above? GNF claim 600 constructs/ week. We will cautiously plan for 400 and 50 weeks / y output == 20,000 /y . So a 5% major user is 1,000/y
Changed:
<
<
I do NMR and I dont think this could help me isotopically label. Any comments? 1) finding the right construct in a eukaryotic screen is likely to be generally useful and be a lead for cell free or prokaryotic expression , and can identify useful complexes, post-translation mods etc.
2) for a few, high profile, cases, the economics of euk expression may be manageable once you know you have high expression. Refs -- LIBRARY:OkhiiDohi09.pdf etc . But realistically we'd have to say this is a research objective once the system is in place. Note that we will have the 1.7 mm TCI 800 probe so small quantities of labeled material are useful.
3) You can think big, e.g. inducible expression of unlabeled materials interacting with injected/ transported labeled materials a la recent Nature papers.
>
>
I do NMR and I dont think this could help me isotopically label. Any comments? 1) finding the right construct in a eukaryotic screen is likely to be generally useful and be a lead for cell free or prokaryotic expression , and can identify useful complexes, post-translation mods etc.
2) for a few, high profile, cases, the economics of euk expression may be manageable once you know you have high expression. Refs -- LIBRARY:OkhiDohi09.pdf etc . But realistically we'd have to say this is a research objective once the system is in place. Note that we will have the 1.7 mm TCI 800 probe so small quantities of labeled material are useful.
3) You can think big, e.g. inducible expression of unlabeled materials interacting with injected/ transported labeled materials a la recent Nature papers.
 
I have another question Please add it to the comment box below

Back to: GetAllFAQs

Revision 2229 Apr 2009 - Main.DavidCowburn

 
Q A
We are assuming that we could not do yeast or Dictyostelium expression - is this correct? It's not clear that the sophisticated procedures available would significantly enhance throughput for yeast. This is something to be explored when the system is established. For Dicty, the issue would presumably be that the current setup is for free cells in suspension, not surface-associated, and that may not be what's needed for your Dicty systems. Finally, there would have to be an operational view of how many different cell systems could be routinely incorporated without cross contamination issues.
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 μ g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioreactors/flasks is a question of cost effectiveness, that users could self determine.
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
how would this facility work- we would be able to provide bacculoviral constructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The initital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm
I think it might be a stretch for me to qualify as a major user, but I can certainly put together a description as an "other user." What's the cross over point? Assuming that cost isn’t a factor (see above) if you can think of 1000 constructs / year to test in your current and hopefully expanded programs going forward -- that’s about the 5% line for a major user. For prokaryotic expression, it’s not unusual for NMR people to make 20-30 constructs per structural target. And this cycle is largely limited by how long it takes to do the structures. For functional studies (especially in any high throughput mode), 50 targets a year sounds very modest implying about 1000 constructs
How did you make up the numbers in the q above? GNF claim 600 constructs/ week. We will cautiously plan for 400 and 50 weeks / y output == 20,000 /y . So a 5% major user is 1,000/y
Changed:
<
<
I do NMR and I dont think this could help me isotopically label. Any comments? 1) finding the right construct in a eukaryotic screen is likely to be generally useful and be a lead for cell free or prokaryotic expression , and can identify useful complexes, post-translation mods etc.
2) for a few, high profile, cases, the economics of euk expression may be manageable once you know you have high expression. Refs -- LIBRARY:AkoiDohi09.pdf etc . But realistically we'd have to say this is a research objective once the system is in place. Note that we will have the 1.7 mm TCI 800 probe so small quantities of labeled material are useful.
3) You can think big, e.g. inducible expression of unlabeled materials interacting with injected/ transported labeled materials a la recent Nature papers.
>
>
I do NMR and I dont think this could help me isotopically label. Any comments? 1) finding the right construct in a eukaryotic screen is likely to be generally useful and be a lead for cell free or prokaryotic expression , and can identify useful complexes, post-translation mods etc.
2) for a few, high profile, cases, the economics of euk expression may be manageable once you know you have high expression. Refs -- LIBRARY:OkhiiDohi09.pdf etc . But realistically we'd have to say this is a research objective once the system is in place. Note that we will have the 1.7 mm TCI 800 probe so small quantities of labeled material are useful.
3) You can think big, e.g. inducible expression of unlabeled materials interacting with injected/ transported labeled materials a la recent Nature papers.
 
I have another question Please add it to the comment box below

Back to: GetAllFAQs

Revision 2129 Apr 2009 - Main.DavidCowburn

 
Q A
We are assuming that we could not do yeast or Dictyostelium expression - is this correct? It's not clear that the sophisticated procedures available would significantly enhance throughput for yeast. This is something to be explored when the system is established. For Dicty, the issue would presumably be that the current setup is for free cells in suspension, not surface-associated, and that may not be what's needed for your Dicty systems. Finally, there would have to be an operational view of how many different cell systems could be routinely incorporated without cross contamination issues.
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 μ g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioreactors/flasks is a question of cost effectiveness, that users could self determine.
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
how would this facility work- we would be able to provide bacculoviral constructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The initital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm
I think it might be a stretch for me to qualify as a major user, but I can certainly put together a description as an "other user." What's the cross over point? Assuming that cost isn’t a factor (see above) if you can think of 1000 constructs / year to test in your current and hopefully expanded programs going forward -- that’s about the 5% line for a major user. For prokaryotic expression, it’s not unusual for NMR people to make 20-30 constructs per structural target. And this cycle is largely limited by how long it takes to do the structures. For functional studies (especially in any high throughput mode), 50 targets a year sounds very modest implying about 1000 constructs
How did you make up the numbers in the q above? GNF claim 600 constructs/ week. We will cautiously plan for 400 and 50 weeks / y output == 20,000 /y . So a 5% major user is 1,000/y
I do NMR and I dont think this could help me isotopically label. Any comments? 1) finding the right construct in a eukaryotic screen is likely to be generally useful and be a lead for cell free or prokaryotic expression , and can identify useful complexes, post-translation mods etc.
2) for a few, high profile, cases, the economics of euk expression may be manageable once you know you have high expression. Refs -- LIBRARY:AkoiDohi09.pdf etc . But realistically we'd have to say this is a research objective once the system is in place. Note that we will have the 1.7 mm TCI 800 probe so small quantities of labeled material are useful.
3) You can think big, e.g. inducible expression of unlabeled materials interacting with injected/ transported labeled materials a la recent Nature papers.
I have another question Please add it to the comment box below

Back to: GetAllFAQs

Revision 2029 Apr 2009 - Main.DavidCowburn

 
Q A
We are assuming that we could not do yeast or Dictyostelium expression - is this correct? It's not clear that the sophisticated procedures available would significantly enhance throughput for yeast. This is something to be explored when the system is established. For Dicty, the issue would presumably be that the current setup is for free cells in suspension, not surface-associated, and that may not be what's needed for your Dicty systems. Finally, there would have to be an operational view of how many different cell systems could be routinely incorporated without cross contamination issues.
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 μ g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioreactors/flasks is a question of cost effectiveness, that users could self determine.
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
how would this facility work- we would be able to provide bacculoviral constructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The initital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm
I think it might be a stretch for me to qualify as a major user, but I can certainly put together a description as an "other user." What's the cross over point? Assuming that cost isn’t a factor (see above) if you can think of 1000 constructs / year to test in your current and hopefully expanded programs going forward -- that’s about the 5% line for a major user. For prokaryotic expression, it’s not unusual for NMR people to make 20-30 constructs per structural target. And this cycle is largely limited by how long it takes to do the structures. For functional studies (especially in any high throughput mode), 50 targets a year sounds very modest implying about 1000 constructs
How did you make up the numbers in the q above? GNF claim 600 constructs/ week. We will cautiously plan for 400 and 50 weeks / y output == 20,000 /y . So a 5% major user is 1,000/y
Changed:
<
<
I do NMR and I dont think this could help me isotopically label. Any comments? 1) finding the right construct in a eukaryotic screen is likely to be generally useful and be a lead for cell free or prokaryotic expression , and can identify useful complexes, post-translation mods etc.
2) for a few, high profile, cases, the economics of euk expression may be manageable once you know you have high expression. Refs -- LIBRARY:AkoiDohi.pdf etc . But realistically we'd have to say this is a research objective once the system is in place. Note that we will have the 1.7 mm TCI 800 probe so small quantities of labeled material are useful.
3) You can think big, e.g. inducible expression of unlabeled materials interacting with injected/ transported labeled materials a la recent Nature papers.
>
>
I do NMR and I dont think this could help me isotopically label. Any comments? 1) finding the right construct in a eukaryotic screen is likely to be generally useful and be a lead for cell free or prokaryotic expression , and can identify useful complexes, post-translation mods etc.
2) for a few, high profile, cases, the economics of euk expression may be manageable once you know you have high expression. Refs -- LIBRARY:AkoiDohi09.pdf etc . But realistically we'd have to say this is a research objective once the system is in place. Note that we will have the 1.7 mm TCI 800 probe so small quantities of labeled material are useful.
3) You can think big, e.g. inducible expression of unlabeled materials interacting with injected/ transported labeled materials a la recent Nature papers.
 
I have another question Please add it to the comment box below

Back to: GetAllFAQs

Revision 1915 Apr 2009 - Main.DavidCowburn

 
Q A
We are assuming that we could not do yeast or Dictyostelium expression - is this correct? It's not clear that the sophisticated procedures available would significantly enhance throughput for yeast. This is something to be explored when the system is established. For Dicty, the issue would presumably be that the current setup is for free cells in suspension, not surface-associated, and that may not be what's needed for your Dicty systems. Finally, there would have to be an operational view of how many different cell systems could be routinely incorporated without cross contamination issues.
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 μ g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioreactors/flasks is a question of cost effectiveness, that users could self determine.
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
how would this facility work- we would be able to provide bacculoviral constructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The initital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm
I think it might be a stretch for me to qualify as a major user, but I can certainly put together a description as an "other user." What's the cross over point? Assuming that cost isn’t a factor (see above) if you can think of 1000 constructs / year to test in your current and hopefully expanded programs going forward -- that’s about the 5% line for a major user. For prokaryotic expression, it’s not unusual for NMR people to make 20-30 constructs per structural target. And this cycle is largely limited by how long it takes to do the structures. For functional studies (especially in any high throughput mode), 50 targets a year sounds very modest implying about 1000 constructs
How did you make up the numbers in the q above? GNF claim 600 constructs/ week. We will cautiously plan for 400 and 50 weeks / y output == 20,000 /y . So a 5% major user is 1,000/y
Added:
>
>
I do NMR and I dont think this could help me isotopically label. Any comments? 1) finding the right construct in a eukaryotic screen is likely to be generally useful and be a lead for cell free or prokaryotic expression , and can identify useful complexes, post-translation mods etc.
2) for a few, high profile, cases, the economics of euk expression may be manageable once you know you have high expression. Refs -- LIBRARY:AkoiDohi.pdf etc . But realistically we'd have to say this is a research objective once the system is in place. Note that we will have the 1.7 mm TCI 800 probe so small quantities of labeled material are useful.
3) You can think big, e.g. inducible expression of unlabeled materials interacting with injected/ transported labeled materials a la recent Nature papers.
 
I have another question Please add it to the comment box below

Back to: GetAllFAQs

Revision 1815 Apr 2009 - Main.DavidCowburn

 
Q A
Added:
>
>
We are assuming that we could not do yeast or Dictyostelium expression - is this correct? It's not clear that the sophisticated procedures available would significantly enhance throughput for yeast. This is something to be explored when the system is established. For Dicty, the issue would presumably be that the current setup is for free cells in suspension, not surface-associated, and that may not be what's needed for your Dicty systems. Finally, there would have to be an operational view of how many different cell systems could be routinely incorporated without cross contamination issues.
 
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 μ g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioreactors/flasks is a question of cost effectiveness, that users could self determine.
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
how would this facility work- we would be able to provide bacculoviral constructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The initital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm
I think it might be a stretch for me to qualify as a major user, but I can certainly put together a description as an "other user." What's the cross over point? Assuming that cost isn’t a factor (see above) if you can think of 1000 constructs / year to test in your current and hopefully expanded programs going forward -- that’s about the 5% line for a major user. For prokaryotic expression, it’s not unusual for NMR people to make 20-30 constructs per structural target. And this cycle is largely limited by how long it takes to do the structures. For functional studies (especially in any high throughput mode), 50 targets a year sounds very modest implying about 1000 constructs
How did you make up the numbers in the q above? GNF claim 600 constructs/ week. We will cautiously plan for 400 and 50 weeks / y output == 20,000 /y . So a 5% major user is 1,000/y
I have another question Please add it to the comment box below

Back to: GetAllFAQs

Revision 1715 Apr 2009 - Main.DavidCowburn

 
Q A
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 μ g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioreactors/flasks is a question of cost effectiveness, that users could self determine.
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
how would this facility work- we would be able to provide bacculoviral constructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The initital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm
I think it might be a stretch for me to qualify as a major user, but I can certainly put together a description as an "other user." What's the cross over point? Assuming that cost isn’t a factor (see above) if you can think of 1000 constructs / year to test in your current and hopefully expanded programs going forward -- that’s about the 5% line for a major user. For prokaryotic expression, it’s not unusual for NMR people to make 20-30 constructs per structural target. And this cycle is largely limited by how long it takes to do the structures. For functional studies (especially in any high throughput mode), 50 targets a year sounds very modest implying about 1000 constructs
How did you make up the numbers in the q above? GNF claim 600 constructs/ week. We will cautiously plan for 400 and 50 weeks / y output == 20,000 /y . So a 5% major user is 1,000/y
I have another question Please add it to the comment box below

Back to: GetAllFAQs

Revision 1614 Apr 2009 - Main.DavidCowburn

 
Q A
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Changed:
<
<
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 μ g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioractors/flasks is a question of cost effectiveness, that users could self determine.
>
>
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 μ g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioreactors/flasks is a question of cost effectiveness, that users could self determine.
 
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
Changed:
<
<
how would this facility work- we would be able to provide bacculviral contructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The intital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
>
>
how would this facility work- we would be able to provide bacculoviral constructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The initital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
 
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm
Changed:
<
<
I think it might be a stretch for me to qualify as a major user, but I can certainly put together a description as an "other user." What's the cross over point? Assuming that cost isn’t a factor (see above) if you can think of 1000 constructs / year to test in your current and hopefully expanded programs going forward -- that’s about the 5% line for a major user. For prokaryotic expression, it’s not unusual for NMR people to make 20-30 constructs per structural target. And this cycle is largely limited by how long it takes to do the structures. For functional studies (especially in any high throughput mode), 50 targets a year sounds very modest inplying about 1000 constructs
>
>
I think it might be a stretch for me to qualify as a major user, but I can certainly put together a description as an "other user." What's the cross over point? Assuming that cost isn’t a factor (see above) if you can think of 1000 constructs / year to test in your current and hopefully expanded programs going forward -- that’s about the 5% line for a major user. For prokaryotic expression, it’s not unusual for NMR people to make 20-30 constructs per structural target. And this cycle is largely limited by how long it takes to do the structures. For functional studies (especially in any high throughput mode), 50 targets a year sounds very modest implying about 1000 constructs
 
How did you make up the numbers in the q above? GNF claim 600 constructs/ week. We will cautiously plan for 400 and 50 weeks / y output == 20,000 /y . So a 5% major user is 1,000/y
Added:
>
>
I have another question Please add it to the comment box below
 
Added:
>
>		Back to: GetAllFAQs
 

Revision 1514 Apr 2009 - Main.DavidCowburn

 
Q A
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 μ g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioractors/flasks is a question of cost effectiveness, that users could self determine.
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
how would this facility work- we would be able to provide bacculviral contructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The intital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm
Changed:
<
<
I think it might be a stretch for me to qualify as a major user, but I can certainly put together a description as an "other user." What's the cross over point? Assuming that cost isn’t a factor (see above) if you can think of 1000 constructs / year to test in your current and hopefully expanded programs going forward -- that’s about the 5% line for a major user. For prokaryotic expression, it’s not unusual for NMR people to make 20-30 constructs per structural target. And this cycle is largely limited by how long it takes to do the structures. For functional studies (especially in any high throughput mode), 50 targets a year sounds very modest inplying about 1000 constructs
>
>
I think it might be a stretch for me to qualify as a major user, but I can certainly put together a description as an "other user." What's the cross over point? Assuming that cost isn’t a factor (see above) if you can think of 1000 constructs / year to test in your current and hopefully expanded programs going forward -- that’s about the 5% line for a major user. For prokaryotic expression, it’s not unusual for NMR people to make 20-30 constructs per structural target. And this cycle is largely limited by how long it takes to do the structures. For functional studies (especially in any high throughput mode), 50 targets a year sounds very modest inplying about 1000 constructs
 
How did you make up the numbers in the q above? GNF claim 600 constructs/ week. We will cautiously plan for 400 and 50 weeks / y output == 20,000 /y . So a 5% major user is 1,000/y

Revision 1414 Apr 2009 - Main.DavidCowburn

 
Q A
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 μ g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioractors/flasks is a question of cost effectiveness, that users could self determine.
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
how would this facility work- we would be able to provide bacculviral contructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The intital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm
Added:
>
>
I think it might be a stretch for me to qualify as a major user, but I can certainly put together a description as an "other user." What's the cross over point? Assuming that cost isn’t a factor (see above) if you can think of 1000 constructs / year to test in your current and hopefully expanded programs going forward -- that’s about the 5% line for a major user. For prokaryotic expression, it’s not unusual for NMR people to make 20-30 constructs per structural target. And this cycle is largely limited by how long it takes to do the structures. For functional studies (especially in any high throughput mode), 50 targets a year sounds very modest inplying about 1000 constructs
How did you make up the numbers in the q above? GNF claim 600 constructs/ week. We will cautiously plan for 400 and 50 weeks / y output == 20,000 /y . So a 5% major user is 1,000/y
 

Revision 1313 Apr 2009 - Main.DavidCowburn

 
Q A
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Changed:
<
<
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 &#micro; g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioractors/flasks is a question of cost effectiveness, that users could self determine.
>
>
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 μ g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioractors/flasks is a question of cost effectiveness, that users could self determine.
 
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
how would this facility work- we would be able to provide bacculviral contructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The intital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm

Revision 1213 Apr 2009 - Main.DavidCowburn

 
Q A
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Changed:
<
<
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 &#mu; g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioractors/flasks is a question of cost effectiveness, that users could self determine.
>
>
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 &#micro; g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioractors/flasks is a question of cost effectiveness, that users could self determine.
 
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
how would this facility work- we would be able to provide bacculviral contructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The intital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm

Revision 1113 Apr 2009 - Main.DavidCowburn

 
Q A
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 &#mu; g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioractors/flasks is a question of cost effectiveness, that users could self determine.
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
Changed:
<
<
how would this facility work- we would be able to provide bacculviral contructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed.
>
>
how would this facility work- we would be able to provide bacculviral contructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed. The intital operation would include a single step slurry separation based on His tag. Final products are produced in 96 well format plates suitable for any further automated purification
 
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm

Revision 1013 Apr 2009 - Main.DavidCowburn

 
Q A
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 &#mu; g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioractors/flasks is a question of cost effectiveness, that users could self determine.
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
Changed:
<
<
how would this facility would work- we would be able to provide bacculviral contructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed.
>
>
how would this facility work- we would be able to provide bacculviral contructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed.
 
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm

Revision 913 Apr 2009 - Main.DavidCowburn

 
Q A
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 &#mu; g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioractors/flasks is a question of cost effectiveness, that users could self determine.
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
how would this facility would work- we would be able to provide bacculviral contructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed.
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
Added:
>
>
this is the first I have heard of your current facility. OK, we are falling down on publicity -- see http://www.nysbc.org/start/index.htm
 

Revision 813 Apr 2009 - Main.DavidCowburn

 
Q A
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Changed:
<
<
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 u g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioractors/flasks is a question of cost effectiveness, that users could self determine.
>
>
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 &#mu; g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioractors/flasks is a question of cost effectiveness, that users could self determine.
 
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
how would this facility would work- we would be able to provide bacculviral contructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed.
Changed:
<
<
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this.
>
>
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this. We are not requesting any financial commitment from you for the HEI application.
 

Revision 713 Apr 2009 - Main.DavidCowburn

 
Q A
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 u g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioractors/flasks is a question of cost effectiveness, that users could self determine.
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
Added:
>
>
how would this facility would work- we would be able to provide bacculviral contructs, and viral titering, Sf9 or Sf21 infections, and two step HPLC would be performed? The initial setup using the GNF procedures would include Sf9. Other systems would be incorporated as the facility progressed.
Do you have any idea what the charges for this might be? On the basis of p 27 of http://www.nysbc.org/HTB/Knuth09.pdf , we estimate about $12 per sample local supply cost (excluding plasmid) . The resource cost with operations and Facility and Administration cost would increase this. We will make every effort to start up the resource at little or no cost to investigators by using additional grants etc., and your additional support may be needed later for this.
 

Revision 611 Apr 2009 - Main.DavidCowburn

 
Q A
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 u g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioractors/flasks is a question of cost effectiveness, that users could self determine.
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.

Revision 511 Apr 2009 - Main.JamesLove

 
Q A
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Changed:
<
<
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 200 - 800 u g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor.
>
>
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 100 - 500 u g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor. However, Scale up in the PEPP versus off line bioractors/flasks is a question of cost effectiveness, that users could self determine.
 
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.

Revision 411 Apr 2009 - Main.DavidCowburn

 
Q A
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 200 - 800 u g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor.
Added:
>
>
Do you prefer I insert the figures in a Word file, or just a PDF file like the sample file? Actually Indesign will be the best to arrange the applications. We would prefer a word2003 file so that minor reformatting can be done. We would accept pdf's if necessary. We may not be able to handle anything else.
 

Revision 311 Apr 2009 - Main.DavidCowburn

 
Q A
Changed:
<
<
Can it work with insect cells. We understand from GNF that Sf9 culture is well established.
>
>
Can it work with insect cells? We understand from GNF that Sf9 culture is well established.
 
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 200 - 800 u g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor.

Revision 211 Apr 2009 - Main.DavidCowburn

Changed:
<
<
Q A
>
>
Qdown A
 
Can it work with insect cells. We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 200 - 800 u g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor.

Revision 111 Apr 2009 - Main.DavidCowburn

 
Q A
Can it work with insect cells. We understand from GNF that Sf9 culture is well established.
Is this more for screening through a lot of different things or for scale up? It's more for screening, but on a pretty large scale and complete to protein purification . We understand that 200 - 800 u g / culture plate are standard and at GNF they will do a batch of four for scale up, but consider that larger scale needs are best done in a larger bioreactor.
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