Advse needed on DNA strand-separation

G G
Fri Nov 11 05:41:30 EST 1994


In article <393h5s$8pl at charm.magnus.acs.ohio-state.edu>, bhuang at magnus.acs.ohio-state.edu (Baohua Huang) says:
>
>Hi, folks. I would like to get some advise on DNA strand separation. I have 1 
>mg of dsDNA of 150 nt long obtained by PCR and would like to separate each 
>strand into ssDNA. Due to the large scale, the avidin-agrose method didn't work
>well (here one strand of the dsDNA contains a bitin at terminus). I intend to
>separate the dsDNA by denaturing PAGE (sequencing gel)because upon
>denaturation, one strand has the extra biotin part which would slow down its
>migration.
>Any suggstions?
>
>Thanks
>
>Baohua
>Ohio State University
I can't quite read your article, but if your ds PCR fragment was produced
with one biotinylated primer, then it is very simple to use Dynals 
streptavidin coated magnetic beads to separate the two strands for sequencing
G Jenkins



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