2 replies to this topic

[katenkak]

Hi everybody,
I am a new area of promoter cloning. My simple question is does it matter from which tissue (of a mouse) I shall purify genomic DNA from or it does not? Moreover, which protocol (commercial kit) is best for such purpose? Many thanks for your time spent answering my questions!

Edited by katenkak, 25 March 2009 - 05:08 AM.

[pcrman]

DNA from any tissue should be fine. For PCR amplification, you don't need any kit, but high fidelity taq polymerase is needed. Because promoter sequences are GC rich, PCR buffer for amplifying GC rich template may help.

[katenkak]

pcrman, on Mar 25 2009, 04:15 PM, said:

DNA from any tissue should be fine. For PCR amplification, you don't need any kit, but high fidelity taq polymerase is needed. Because promoter sequences are GC rich, PCR buffer for amplifying GC rich template may help.

Hi!
Thanks a lot for answer and advice on PCR buffer. Is there a protocol for genomic DNA purification that you know works well? I would be thankful if you share one with me.