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I've being using this lysis buffer for fungal DNA extraction,lysis buffer - 400mM TRis-HCl(pH8.0), 60mM EDTA(pH8.0), 150mM NaCl, 1% SDS and containing 40microgram/ml RnaseA.

It is keep giving very light bands on 1% agarose gel and shear appearence. After extraction I have to follow PCR and sequencing. If there are inhibitors, how can I get rid of those ?

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    $\begingroup$ Do you Fungi contain pigment, especially melanin? This is a potent inhibitor for PCR. If not, you could start by trying a phenol/chloroform extraction. $\endgroup$ – Chris Oct 22 '15 at 6:23
  • $\begingroup$ yeah they have pigments,one sample was a penicillium, i did PCR for these DNA extractions (didn't make any changes to the lysis buffer) but I got good bands after PCR,my primers were ITS1 and ITS4. But I didn't get PCR results for extractions I did more than 4 days ago. thank you for the answer. $\endgroup$ – k.m Nov 7 '15 at 9:08

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