BAC DNA purification

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BAC DNA purification

Post by kirupha » Wed Mar 07, 2018 8:54 pm

Hi all,

I am trying to purify BAC DNA (106 kb) using Qiagen large construct kit. I am able to purify 5-10 ug of DNA but when I run on 0.8 % agarose gel I see a band near the well. After restriction digestion also I see this band but I can detect it with southern. But we need a clean prep to give it for pronuclear injection into mouse embryo. I tried phenol/chloroform to remove proteins but this seems to be genomic DNA. I have attached agarose gel picture (0.8 % gel) of undigested BAC DNA. Kindly help me to solve this problem

agarose gel.jpg

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