synthesising ds oligo's using klenow poly.

GRGGTA at PATERSON.CHRISTIE-HOSPITAL.MANCHESTER.AC.UK GRGGTA at PATERSON.CHRISTIE-HOSPITAL.MANCHESTER.AC.UK
Mon Jul 26 10:56:00 EST 1993


Len Adleman writes:-
Dear Colleagues,

I have 40 nmol of a 58 nucleotide oligo. I also have 40 nmol of an 18 
nucleotide primer complementary to the 3' end of the oligo. I wish to
double strand the oligo, but have not been able to find a good protocol.
Is there a good one? How much can I do in one reaction?

-Thanks
-Len Adleman
-USC
-adleman at pollux.usc.edu

A method I have used for this is referenced in Chodosh et al, Molecular
and Cellular Biology vol6 p4723-4733. I made nanogramme quantities of
ds oligo using this method for use as radioactive probe containing 
deoxybromidine, but I see no reason why it should not work better using
'regular' dNTP's.
Graham Atherton



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