Annealing of complementary s.s. DNA's
Mon Feb 13 18:52:44 EST 1995
In article <amatos-1302951347370001 at bib7.bc.edu> amatos at hermes.bc.edu (Stephen Amato) writes:
> I have recently ordered some complementary oligo's from BioServe
>Biotechnologies. These oligo's have 5' overhangs and I wish to label the
>annealed products using Klenow and alpha radiolabelled deoxynucleotides.
>I have been having some trouble labelling them and I want to make sure the
>single stranded precursors are actually annealed. Does anyone know of a
>method or of a reference that I could look up that would contain a
>protocol for actually following the annealing, maybe by absorbance at
>260nm? I suspect one of think my other reactants may not be O.K., but I
>just want to make sure that my oligo's are indeed annealed.
>amatos at hermes.bc.edu
Run a gel. Gel conc will depend on the size of the oligos. the ds should have a
slower migration rate through the gel than the ss. High conc agaroses are best
since the molecules run tru to size and are not affected by GC content.
Uni of Sydney
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