old degraded samples. DNA extraction & PCR - (Dec/23/2004 )

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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Hi

i am havingthe same problem (old fish samples) try to optimise my extraction. which protocol worked best for you??

greetings

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Why not try boiling in chelex-100. It's been used for DNA extraction of fixed or non-fixed tissue, typically for forensic or museum samples. If you're only interested in running PCR it's probably the way to go.

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Hi,

I've never worked with such old fish samples, but I used to extract DNA from fresh tissues and found that fish DNA seems to degrade much faster than DNA from other sources. This might prove problematic for you. If your samples weren't preserved soon after death of the fish in 1910, getting usable DNA now might be impossible, but it's worth trying what KPDE suggests, or other methods developed for extracting DNA from old material. Good luck, and let us know the results.

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