Hello,

I've already done this with Qiagen Tissue extraction kit, following their very simple protocol, and it worked perfect. I used one leg, but it was almost too much for some samples. When dealing with large species, I would recommend to use one little piece of leg only. Good luck !

Dear Vincent,

great.  Can you tell me for which bee species you used it? And how old were the specimens? Is there anything I have to be particularly careful about?

Thanks a lot for your help.

Megachile sp., Bombus sp. and Osmia sp.

It was a series of tests for barcoding, but I tried to amplify 28S and it worked well too. Most specimens were only 2 years old, so I don't know if really old bees will work that well.

Some colleagues working with Homoptera tried with many old dried specimens (at least several decades old) and the results were not as good, but we don't know if some treatment degraded their DNA at some point. It depends a lot on the conditions of storage and preservation.

Thanks. I don't really know how the specimens were collected and stored. What do you mean with 'not as good'- it did not work at all or you just got low quality sequences? Thanks for your help again.

Well most of the time amplification "worked" in that we got PCR products, but with high contamination rates (Homo sapiens, Fungi, bacteria, even plant sequences...). If you don't know how they were killed and stored, best is to give a try with some specimens.

For bees in the genus Melissodes, I had trouble getting DNA from specimens older than about 10 to 15 years. I also just used the Qiagen tissue extraction kits. For older specimens, I have heard that adding DTT helps, but I have never tried it. I used a middle leg from each specimen and crunched it up with smooth forceps to expose the muscle tissue.  The younger the better and try to avoid anything that has been killed by Ethyl acetate or has been stored in less than 95% EtOH.