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old degraded samples. DNA extraction & PCR


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#1 GT(N)T

GT(N)T

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Posted 23 December 2004 - 05:29 AM

I have muscle samples of fish that were collected in 1910 and stored in a museum. The curator isn't sure if they were ever fixed in formalin, but they were in ethanol when I pulled the samples.

So far I tried using PureGene kits and protocols to extract DNA. The results are indifferent. The last spin leaves a faint streak on the tube that appears to be DNA. Electrophoresis shows a broad smear.

PCR using both universal and congeneric mtDNA primers have been unsuccessful. We were able to amplify a microsat band of the right size in one animal using congeneric primers and TD50 protocol.

Any suggestions to improve recoveries in old degarded tissue or in PCR of the recovered DNA?

Thanks y'all




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