I need to purify some genomic DNA from a human skin punch biopsy. In the past, I have done this with the Qiagen DNEasy Blood and Tissue kit. This yields good quality DNA for routine PCRs, however, I now need to amplify a 14kb fragment from this DNA. The PCR works if the template DNA is unfragmented, but the DNEasy DNA does not work. Is there a better way to purify DNA from human skin so that it doesn't end up fragmented?
I think the genomic DNA prep would still be containing fragment with more than 14 kb size and you can get amplification. I hope if you have already tried a PCR, it was using a high fidelity DNA polymerase such as Q5 which is recommended for amplicons of upto 20 kb size. Other low processivity polymerase might not be able to function.
I am using Primestar GXL from Takara, the 14kb PCR works fine on other templates.
As I can see in it's manual, it's a high fidelity polymerase good for the purpose, so may be there is some issue with the template.
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