DNase I digestion - check if it worked?? (Apr/27/2009 )
i have to add a DNase I digestion step following my RNA extraction. how will i know if this digestion worked or not? agarose gel?
maybe you have some recommendations/suggestions!
just add an RTminus control. Do duplicate reverse transcription reactions with your samples but leave out the RT enzyme (=RTminus control) in one (just add water instead). Then do PCR with both. If your primers detect gDNA and there was some gDNA left in your RNA prep you will get a signal in the RTminus control.
tea-test on Apr 27 2009, 07:51 AM said:
right, i thought so complicated!
You don't need to do duplicate reverse transcription without the enzyme.
Just take some of your RNA sample as template for standard PCR!
Yes, you should take an aliquot and run the gel. You might see all your RNA's gone (over digest) or you might see a large DNA smear (under digest).