Construction of BAC library - (Feb/11/2009 )

Hi

I would like to construct a BAC library for a fish species, but I have no experience in this technique. Does anyone have any kit or protocol to recommend? It would be highly appreciated.

Hello

I don't do it myself but know of many papers where they use pBeloBAC11, digest it with some site with cuts the genomic DNA at a relatively less frequency ( to ensure big fragments), digest the DNA and ligate. Ideally, I would just digest the genomic DNA, pool multiple batches and precipitate. The vector, I would digest, CIP, pool and precipitate. Then resuspend both and ligate.

This should work but I really don't have experience doing this and would highy recomment that you see some sequencing paper and see how they have done it. I know that BAC clones exist for Mtb DNA.

Hope it helps.

TC

cathrief on Feb 11 2009, 08:37 PM said: