purifying out one strand of PCR product?

Eirikur Sigurdsson 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

>Johnny D

>-- 
>John Dixon                     Lab 44 (223) 334131
>Wellcome/CRC Institute         Fax 44 (223) 334134
>Dept Genetics
>Cambridge University    
>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.

Eirikur Sigurðsson
Laboratory of Molecular Genetics
Institute of Biology
The University of Iceland
Reykjavik		email:  eiriksig at rhi.hi.is



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