Salt-free DNA precipitation?
Koen De Smet
Thu Nov 9 09:07:35 EST 1995
diabetes at lrz.uni-muenchen.de (Wolfgang Schechinger) wrote:
>I'm new to PCR, so let me ask a question, please: Is this a common
>protocol or is it home-brew? If you have one, please give me
>Koen De Smet <kds30> wrote:
>>I routinely precipitate PCR products by adding 1/10 V of 3M sodiumacetate pH 5.2 and 1V of 30% PEG-6000.
>>Leave for 15 minutes at room temperature and spin. Wash with 70% ethanol.
>>This precipitates the PCR product, but not the oligos, so the DNA can be used for sequencing.
>>Koen De Smet
I was given this method by somebody else, who used it for sequencing PCR products using an automated sequencer (I don't know which model). I believe he got this method from the manual that came with the machine. It works fine and gives good recoveries, except that I sometimes lost all DNA. But maybe that was due to careless washing...
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