Prep protocol for BAC clones - (Aug/21/2006 )
I used to make pools of BAC clones by doing midi preps of 96 clones plated together in one Petri dish. The protocol used for that was the "basic" prep with lysis buffer containing NaOH 200 mM SDS 1% and neutralizing solution of potassium acetate 3 M. The results were generally good with variable yields. Now I intend to make a prep from one BAC clone to use it to construct a shotgun library of this clone. Some people told me to use Qiagen column to purify the extraction. Do you all think it is necessary to have a high quality DNA to make a library?
Thanks in advance!
High quality DNA is good for a library. Its debateable whether or not Qiagen kits give you good quality DNA. But their kits will give you sufficient quality to make your shotgun library.