Bolitophagus Bug Bath is a Bust: Gilbert (2007) Extraction Protocol Fails in First Attempt
As I mentioned in this entry, I ran the Gilbert (2007) protocol for extracting DNA from dried beetles. I got no result at all when I attempted to precipitate the DNA from two Bolitophagus reticulans specimens collected in 1992. I simultaneously attempted a destructive protocol on two other specimens from the same collection. Both of these attempts resulted in the precipitation of what I am assuming for the moment is DNA.
Tomorrow, I'll attempt to amplify the precipitant from the destructive extractions. I'm also going to attempt the non-destructive extraction once again, this time using the freshest specimens available - two lobster cockroaches (Nauphoeta cinerea) which are still alive at the moment. While it is possible I made some error in following Gilbert's protocol, it may be that my Bolitophagus specimens were too old or too poorly preserved and that this protocol is limited to the extent that there need be a relatively large amount of intact DNA, even in a dried specimen.
If I can amplify and sequence from both the destructively extracted Bolitophagus and from the very fresh Nauphoeta, I'll have better data regarding the limits to the usefulness of the protocol itself.
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
