I'm trying to extract genomic DNA with organic extraction from cell lysates in order to quantify the amount of DNA in the sample. However, I'm getting inconsistent readings that I think are due to RNA contamination in the lysates.
I have many samples so would like to avoid using RNAse, is there a way to use alkaline lysis to destroy the RNA without damaging the DNA?
Hi Ahmad
To eliminate RNA, you may use an RNase treatment protocol. This will degrade all RNA molecule and wont affect DNA quantity, although still you may have some problems with reading! The best recommendation is to use an extraction procedure which can extract only DNA.
Good luck
You may not use alkaline hydrolysis for RNA, this will destroy your DNA also. RNAse is best way as Ali mentioned. No short cut for good results
RNA is much more susceptible to hydrolysis at high pH than DNA is, so you can inactivate RNA that way. However it won't solve your problem, nor will RNase treatment. Both of those will break down longer RNA polymers but leave the bases intact which will still absorb at 260nm. (Although a subsequent ethanol precipitation step would help). Your choices are to measure your DNA concentration in a different way or to better purify your DNA so as to remove the RNA.
I should add that you inconsistent readings might be from RNA, but they might also be from residual chemicals since you are doing organic extractions. For example any traces of phenol will give very high 260 readings.
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