February 04, 2008

Gilbert (2007) DNA Extraction: Specimens Too Poorly Preserved

ResearchBlogging.orgAs a follow-up to my entry of two days ago, it looks like my failure to extract DNA from the dried Bolitophagus reticulans specimens may be due to the age and/or poor preservation of the specimens and not to any defect with the protocol itself.

Today, I attempted to amplify genomic DNA from the specimens using a well-tested, standard protocol. I used several primer sets, including LR0R-LR7 (lsu), ITS1-ITS4, and a couple of custom-designed primers meant to capture chitinase genes. I got nothing no PCR products from either of two specimens collected in 1992:

My next step will be to test Gilbert's protocol against the standard using fresh specimens, as mentioned.

Gilbert, M.T., Moore, W., Melchior, L., Worobey, M., Hofreiter, M. (2007). DNA Extraction from Dry Museum Beetles without Conferring External Morphological Damage. PLoS ONE, 2(3), e272. DOI: 10.1371/journal.pone.0000272

(PS: Sorry for the double-post; I'm attempting to get my feed to work with ResearchBlogging.org's aggregator. I needed to repost in order to try to fix a problem that's been preventing it from working correctly).

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