Hello,
I am trying to generate ssDNA from an RNA template via RTPCR and consecutive digestion of the remaining/duplexed RNA by RNAse H.
The first part is easy, but if I run my ssDNA on a gel I get 2 bands instead of one (see picture attached). First lane is DNA precipitated by phenol/chloroform and the second lane is crude RNAse H digested product. The heavier band has the correct size of ~900bp. What is the second band?
Longer digestion didn't help.
Also, (not shown here) the RNA template as well as the cDNA prior to RNASe H digestion give clean bands at 900bp.
This is a non-denaturing standard agarose gel.
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