RNase in RNA - QC for RNase in RNA (Aug/18/2005 )

Hi,

I wanted to check for any Rnases in my RNA fragments after transcription reaction and I did this by taking portion of my RNA sample and mixing with ligase buffer and incubating it for about an hour at 37 degrees celsius....

After an hour, I took the RNA sample with ligase buffer and ran it on denaturing gel along with a control sample (same RNA, no ligase buffer or incubation at 37). THe control was ok, but the sample with ligase buffer showed degradation...I denatured both samples at 95 degrees for few minutes before loading onto gel....Should I have not done that with the RNA in ligase buffer and why?

What was in the ligase buffer? At 37C cations such as Mg will chemically degrade RNA

I use Ambions RNaseAlert kit for checking for RNAse contamination, it's great

RNA doesn't like high temperatures in the presence of divalent cations (like Mg2+).

Daniel

DNA sequencing primers

RNA doesn't like high temperatures in the presence of divalent cations (like Mg2+).

Daniel

DNA sequencing primers