Good day, we have a problem with recent RNA that we obtained from a plant, we suspect that it might have PCR inhibitors. One of the possible solutions that we thought was to do isopropanol precipitation of the RNA, but we are wondering if it is possible to treat it with DEPC to make it free of RNAses. Has anyone tried this or has a different solution?
To deal with polyphenols and other PCR inhibitors (nucleases) in plant DNA-RNA extraction, we used to add the β-Mercaptoethanol (2-ME):
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Some pcr inhibitors bind Mg making the Taq useless so a titration of MG ions from 1.5mM to 5mM sometimes helps. Another way of dealing with inhibitors is to titrate the amount of template downwards. As pcr is so productive often using less template also means having less inhibition and once the pcr can work the shortage of template initially is made up for in the amplification cycles
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