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-   -   BAC library screening using PCR (http://www.molecularstation.com/forum/molecular-cloning-forum/84622-bac-library-screening-using-pcr.html)

Irrateelevatorguy 02-08-2012 02:57 PM

BAC library screening using PCR
 
Hello,

I am currently working on a project where we have received a BAC library with 100 clones pooled per well. My boss has said we will screen using PCR. We will do an initial screen with PCR, then for the wells which received a postitve reaction we will dilute then 1/96 and PCR again. We will then sequences using 454 the diluted positive wells. I cannot find any material online about this, and I am not sure if my supervisor knows an exact protocol. I am slightly confused about this whole method, is there anyone who can further explain it to me or provide me with a reference on this???

Any input is greatly appreciated, thank you.

Irrateelevatorguy 02-08-2012 03:08 PM

Re: BAC library screening using PCR
 
We are not using superpooling and 3D plates and PCR etc. because we only have plates with 100 clones pooled per well.


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