it would also work if you put it in a heating block and follow the previous
directions. actually i usually don't cool to 4C just to room temp and they
have worked fine for restriction digests and subsequent ligations.
"Scott Coutts" <scott.coutts at med.monash.edu.au> wrote in message
news:3DA1798B.5030300 at med.monash.edu.au...
> I'm not exactly sure what you're asking...
>> Do you mean that you have two single-stranded DNA molecules that you
> need to anneal together, to make a double stranded segment which you
> then want to ligate into a vector? If so, just heat them up to 90
> degrees C, leave to cool to room temperature and then put them on ice.
> They'll anneal if they're complimentary.
>> Sorry if I've completely missed the point of your question!
>> Isaac Xin Pei wrote:
> > Hi, Molecular Biologists,
> > I have designed a long sequence tag about 70mers; after obtaining the
> > primers, now I need to anneal them, and ligate them to my vector. Does
> > anyone has a protocol for annealing the long single-complentary strand
> > for these works? Thanks a lot!
> > Best, Isaac