Hi all,
I am attempting to get enough (and good enough) DNA extracted from dried adult butterflies to do sequence capture. I've been told I need at least 1 microgram of DNA for guaranteed results, which is a bar I have not yet met (0.952 micrograms is the most I've gotten, to date).
I have been mechanically grinding a single leg per specimen, and then using overnight incubation with proteinase K, followed by phenol chloroform extraction (the commercial Omniprep kit), according to the standard procedure (except that I'm using Glycoblue instead of the glycogen that comes with the kit).
Does anyone have recommendations for tweaks this procedure, or other procedures that may increase yeild?
Otherwise, when do you say enough's enough (for quality and quantity) and send it off and hope for the best?
Thanks for your thoughts.
I am attempting to get enough (and good enough) DNA extracted from dried adult butterflies to do sequence capture. I've been told I need at least 1 microgram of DNA for guaranteed results, which is a bar I have not yet met (0.952 micrograms is the most I've gotten, to date).
I have been mechanically grinding a single leg per specimen, and then using overnight incubation with proteinase K, followed by phenol chloroform extraction (the commercial Omniprep kit), according to the standard procedure (except that I'm using Glycoblue instead of the glycogen that comes with the kit).
Does anyone have recommendations for tweaks this procedure, or other procedures that may increase yeild?
Otherwise, when do you say enough's enough (for quality and quantity) and send it off and hope for the best?
Thanks for your thoughts.
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