Hello,
We recently obtained some insect samples (Asphondylia) collected a few months ago by collaborators. The individuals were freeze dried, placed into pure ethanol, and shipped to us on dry ice.
I am having trouble getting a good DNA concentration from them (~50ul, 0.5ng/ul). I have tried the Qiagen DNeasy blood and tissue kit, phenol chloroform, and the Lucigen MasterPure Complete DNA & RNA Purification kit. I have tried varying incubation times (up to 17 hours) without any increase in DNA yield. I've been using the Quiagen Tissuelyser with tungsten beads to ensure thorough homogenization.
Does anybody have any protocols or suggestions on how we can get a higher yield? They are quite small insects (like mosquitoes), but I need a minimum concentration of about 5ng/ul in 150ul!
Thanks!
Hi, I'll try to rehydrate my insects and add some distilled water (1/1 volume) in my tubes before the extraction to dilute ethanol. Then discard supernatant, and add water again and again.
Good luck :)
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