I have been using Omega and QIAGEN DNA tissue and blood extraction kits to extract the DNA of mud crabs (Scylla spp.) gut content, but both did not give good results. I have tried 8 different (8 times trials) combinations on incubation time, sample (gut content) quantity (ranged from 3 to 30 mg, as suggested by the kits' guidelines), sample pre-treatment (diluting it with distilled H2O to remove any excessive remaining preservative (96% of ethanol), mincing the sample, and making sure any extraction reagents needed are treated according to the kits' guidelines (dissolving any precipitation occurred on the reagents). Based on the nanodrop, i only received 9-28 nanograms of DNA/ ul, with low purity (A260/A280) ranging from 1.39-1.47). There was no band shown on the electrophoresis gel either.
My biggest suspicion is still on the sample amount. How much (mg) of gut content did you use? (would you mind to share the photograph as well) did you use any pre-treatment to the tissue before diluting it with Proteinase-K? I was thinking of using liquid Nitrogen to speed-mince the tissue prior to the Proteinase-K dilution. I look forward to hearing your opinion.
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