aaptor problems

Mr JN Goulding jgouldin at hgmp.mrc.ac.uk
Tue Jul 25 05:05:51 EST 2000


Hi. I have been having a problem with annealing together two oligos to
produce an adaptor. Basically, during the annealing stage the singl;e
stranded ends of the adaptor are degrading, meaniung that some does nopt
ligate to my DNA fragments and is being carried through into my PCR, and
acting as a primer, causing background problems. I've devised a methof to
get round this, but I would like to know why it is happening, and if
anyone else has had these problems. The method I use for annealing the
adaptors (heat then a slow cool) I've used previously with no
problems. I've tried different manufacturors of oligos to no avail, adn I
know that other groups elsewhere we collaborate with using the sam,e
methods have the same problems. Any ideas or suggestions would be
appreciated. Cheers everyone, have fun.

Jon






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