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Low copy vector for library generation


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#1 pDNA

pDNA

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Posted 10 April 2013 - 08:10 AM

Dear people,

i'm currently trying to generate a sigma-factor library using pHSG576-derivative (SC101ori, low copy) as backbone. I'm trying to get a library size of 10E5-10E6 ...but i'm far away from that currently. I have the feeling that the backbone is one of the major problems.

Therefore, i would like to ask if anybody has suggestions for a low copy plasmid backbone that has been used excessively for library generation?
I was thinking about pBELOBac11? Any other suggestions?

In addition, does anyone has a good references (book, journal articles) that deal with library generation in E. coli?

I very much appreciate your help!

Many thanks in advance!

Best regards,
p

#2 phage434

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Posted 11 April 2013 - 02:44 PM

Very old school is lambda libraries. These have the advantage of very high transformation efficiency, and rapid replication. p15A origins are not quite as low copy as pSC101, and are pretty user-friendly.

#3 pDNA

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Posted 19 April 2013 - 01:17 AM

many thanks for your reply!
Does anyone know how stable the pBeloBAC11 is ...can it be maintained without antibiotic selection pressure (does it include a toxin/antitoxin system?)?

Many thanks in advance!

Best regards,
p




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