psoralen protocol

Balaji Kannan balajik at newton.berkeley.edu
Tue Jul 24 19:16:51 EST 2001


hi there-
    i am looking at ligation of oligonucleotides without using DNA ligase-
using psoralen. the idea is that the strands that are to be ligated are
first hybridised to a template that's complementary to both strands. one of
the oligos to be hybridised has psoralen at it's 5' end attached to base T.
the other strand is properly structured so that psoralen can intercalate
when UV is shined. the only problem seems to be that the thing doesnt work
at all. i've tried irradiation using long UV for anywhere between 2 minutes
and 4 days and all results are negative.

    could anyone please give me a a pointer to where i can look for a
protocol that could help me do the thing i'm looking at, including how is
should shine the UV light (whether directly or thru tubes etc).

    i'd be grateful for any help.

    thanks in advance

    balaji





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