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Removing any traces of DNA

John Dixon jpcd0 at mole.bio.cam.ac.uk
Thu Dec 21 06:57:26 EST 1995

In article <aquilla.1169708309O at emba-news.emba.uvm.edu>,
aquilla at salus.med.uvm.edu (Tracy Aquilla) wrote:

> >On Thu, 14 Dec 1995, Jean Peluso wrote:
> >
> >> I would like to know how I can remove completely DNA from a preparation
> >> of in vitro synthetised RNA. I have already tried the DNAse I recommended
> >> by Invitrogen (T3 Transcription Kit), but when I perform RT-PCR on the
> >> RNA  preparation I unmask traces of DNA, even by using more DNAse and


> has RT activity? I routinely use RNAse-free DNAse I from Ambion to eliminate
> contaminating DNA from my RNA preps and haven't had any problems eliminating
> the contaminating DNA. This enzyme is so clean you can do a several-fold
> over-digest without affecting the RNA. Have you tested the effectiveness of
> your DNAse solution?
>     Tracy

How do you guys then get rid of the DNAse1 prior to cDNA synthesis? If a
phenol chloroform extraction is used, must care be taken to use RNA phenol
ie low pH phenol to avoid loss of the RNA to the wrong phase?

John Dixon                                        Lab 44 (1223) 334131
Wellcome/CRC Institute                            Fax 44 (1223) 334134
Department of Genetics
Cambridge University    
United Kingdom                           e-m: jpcd0 at mole.bio.cam.ac.uk

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