Gilbert (2007) DNA Extraction: Specimens Too Poorly Preserved
As 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.
Reference:
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).
