I have red poultry mite samples which have been stored in ethanol. Now I need to perform RNA and DNA extraction followed by shotgun metagenomic sequencing. I would like to know if there is a way to successfully extract the nucleic acids so that I have no inhibition or problems during the sequencing run. You opinion and experience is highly appreciated.
You may refer chapter 6 of "Molecular Cloning a laboratory manual" 4th edition by Michael R. Green and Joseph Sambrook. The chapter contains impressive details about the different extraction protocols and methods.
ISBN 978-1-936113-41-5.
or you may refer to this book.
RNA Isolation and Characterization Protocols by Ralph Rapley, David L. Manning.
Hope this helps.
Pretty much all RNA/DNA extraction kits (Qiagen, Zymo, or even homogenization followed by etoh precip) will remove inhibitors. You may worry more about the degradation of nucleic acid especially if it was stored in ethanol for an extended period of time - you can check that on a Bioanalyzer.
Péter Gyarmati thank you for answer! I had a bad experience when I performed PCR on samples that had previously tested positive for some viruses. After this PCR the samples (original samples, not nucleic acids) were stored in ethanol for some time and when I performed PCR again it was negative. Do you have any idea why this happened? As for the degradation, that is not a problem, the samples are not stored for an extended period of time. Malith Arambage thank you for your asnwer, I'll check it out.
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