Annealing of two complementary circular SS DNAs

Lloyd Graham graham at dbe.csiro.au
Thu Aug 15 21:01:29 EST 1996


A procedure I am conducting will generate two closed circular
single-stranded DNAs which are perfectly complementary to each other. I
would like to anneal them together to generate a double-stranded DNA (a
3.6kb plasmid). Is it simply a matter of heating to 95'C and allowing them
to cool slowly, or will their circular nature complicate things?

Ultimately I just want to transform E.coli with the resulting plasmid.
Perhaps it's not even necessary for me to do an annealing step - does
anybody know if transforming with single-stranded plasmid DNA is
effective? 

Many thanks,

Lloyd Graham
CSIRO Div. Biomol. Eng., Australia



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