How to eliminate gDNA from RT-PCR?
Dr. Duncan Clark
duncan at genesys.demon.co.uk
Wed Jul 23 07:47:13 EST 1997
In article <5r2uv7$2pd at cardinal2.Stanford.EDU>, John Ladasky
<ladasky at leland.Stanford.EDU> writes
>1) Use a different method of mRNA isolation. Does anyone know
> that the method of mRNA preparation that they use is particular-
> ly good at removing gDNA?
> 2) Treat the mRNA with DNAse. A colleague in my lab has tried this
> without success. She thinks that most DNAse preps have contam-
> inating RNAse activity (and I agree that this is likely).
When you get some RNase free DNase, try digesting in a manganese buffer
rather than a magnesium buffer - Biotechniques June 97 p1128-1134.
The problem with being on the cutting edge is that you occasionally get
sliced from time to time....
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