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.

Duncan

-- 
The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
DNAmp Ltd.
TEl/FAX 01252376288
http://www.dnamp.com
http://www.genesys.demon.co.uk



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