T4 DNA Ligase: Oligo to Sticky End

Paul R. Rohde P.Rohde at mailbox.uq.edu.au
Tue Nov 25 03:10:30 EST 1997


T4 DNA Ligase joins blunt ended or cohesive-ended double stranded
molecules but does not join single stranded molecules.

My question:  Will T4 DNA Ligase join to a four base sticky end,a
complementary oligonucleotide?  Yes, the oligo is single stranded,
but the actual ligation site (or nick) is actually double stranded.
This should work shouldn't it?  The oligo has been kinased to the 5'
end.



                         5'-gatcnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn...
     3'-NNNNNNNNNNNNNNNNNNNNCTAGnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn...
                               ^^
                             "nick"


        (Oligo Capitals, ds Sticky End DNA lower case)



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