Separating ssDNA from dsDNA (asymmetric PCR) ???

William Alexander ALEXANDERW at cber.cber.fda.gov
Wed Oct 11 03:12:42 EST 1995


In Article <1995Oct10.171838.17067 at newsserver.rrzn.uni-hannover.de>, bmay
<ndxdbmay at rrzn.uni-hannover.de> wrote:
>Does anybody out there know a simple way to separate ssDNA from dsDNA 
>(~250bp) and other compounds in the PCR mixture after asymmetric PCR ? 
>I want pure ssDNA for sequencing without expensive and time consuming 
>procedures as gel-cutting and gel-extraction-kits and so on.
>
>Thanx a lot in advance
>
>Bernhard Mayr
>Medizinische Hochschule Hannover
>Dept. Clin. Endocrinology
>Hannover, Germany

Is this separation really necessary?  Just ethanol precipitate, wash,
resuspend and sequence.  Double stranded plasmids can be sequenced without
purifying the strands, why not this small DNA?  Is there some redundant
sequence that causes trouble?

Bill Alexander
alexanderw at cber.cber.fda.gov



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