When we are trying to sequence COI of bees and parasitoid wasps, we always collected legs and stored them in pure alcohol. However, we failed to get COI from many samples with good DNA. If you have some experience on these group or even other animals, please kindly let me know.
Can you please provide us with informations about primers that you are using? I am working on some parasitic wasps and I had just I few failure in isolation. I use very old primers LCO1490 (5’-GGTCAACAAATCATAAAGATATTGG-3’) and HCO2198 (5’- TAAACTTCAGGCTGACCAAAAAATCA-3’) (Folmer et al. 1994) whic work almous perfectly for very diverse organisams.
I hope this will help.
If there are not specific primers for bees you could try to amplify the barcode region with "universal" primers as Andjeijko refered. You could use these primers too:
http://www.biomedcentral.com/1472-6785/13/34
Andjeljko Petrović, Im using those but may be yo can try using some of the ones reported by Simon et al 2006. In the Supplements you can search the group you are working with...
hope it was useful
I have worked with Ephemeroptera, and now I'm working with Chironomidae, when i used just legs the amount of Dna was usually low and we didn't get good results, I usually use the whole insect, in some cases without gut. With Chironomidae problems Amplifying COI are less frequent, may be the primers are not the ones???
I have compared tissue extraction used for insect (not published) in terms of DNA concentration and A260/A280 and A260/A230 ratio. I found that the optimal protocol was to extract DNA from head + thorax. This test was based on large invertebrates panel.
I used also a coktail of LepF1 LepR1 with LCOI HCO2, and it works in 85% based on 180 invertebrates samples.
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