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Hello,

I am currently completing my PhD, which looks into how altering the belowground mycorrhizal community has the potential to influence pollinator communities in strawberry plants. Over the past two years I have collected a number of pollinators visiting my plants, and I am now in the process of identifying them all. I am confident taxonomically identifying hymenoptera and hoverflies, however I have been advised that taxonomic ID of the remainder of the Diptera is rather difficult, and that I am better off identifying them through barcoding. So far I have tested a NaOH extraction method, along with a separate Proteinase K extraction method, however both haven't been very successful, and I am seeing a success rate of less than 40%. I think that this may be due to the fact that the some of the samples were pinned this time last year, and some over the last month, and in turn the fact that they have now dried, which is causing these issues.

I was wondering if anyone has experience in barcoding museum specimens of Diptera/insects that have been dried, and would be able to provide any advice on the methods that you use? In an ideal world I would prefer to just extract a leg from each sample if possible, in order to keep the remainder of the sample for a reference collection.

Thank you very much ahead of time, and I hope you have a nice evening!