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.
0
1 reply to this topic
#2
T C
-
- Active Members
-
- 277 posts
Veteran
0
Neutral
Neutral
Posted 11 February 2009 - 11:25 PM
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
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:
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.
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.
