purifying out one strand of PCR product?
eiriksig at rhi.hi.is
Tue Jun 6 12:52:44 EST 1995
In <jpcd0-0606951337540001 at macr1-4.welc.cam.ac.uk> jpcd0 at mole.bio.cam.ac.uk (John Dixon) writes:
>Hi, can anyone tell me what is the easiest and/or most efficient way to
>purify one strand of a PCR product from the other?
>The product of the PCR will be about 100bps but I wish to use one strand
>as a primer for a reverse transcription reaction and I do not want
>contamination from its complementary strand.
>Is it possible to selectively degrade one or other strand?
>Thanks in advance
>John Dixon Lab 44 (223) 334131
>Wellcome/CRC Institute Fax 44 (223) 334134
>United Kingdom e-m: jpcd0 at mole.bio.cam.ac.uk
I think the easiest way is to use biotinylated primers. After denaturating
the DNA you can isolate the biotinylated strand with streptovidin coated
magnetic beads. I have used this method for isolating single stranded DNA for
sequencing and it works very well.
Laboratory of Molecular Genetics
Institute of Biology
The University of Iceland
Reykjavik email: eiriksig at rhi.hi.is
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