Blunt end liagation problems

mbro at seqnet.dl.ac.uk mbro at seqnet.dl.ac.uk
Thu Oct 19 03:20:28 EST 1995


Your problem may stem from the fact that you are using Klenow to blunt
end your DNA. Like Taq, but to a much lesser extent, this enzyme also
adds additional nucleotides to the 3' end to a significant poplulation of
of your DNA.  The best enzyme for getting blunt ends are T4 DNA polymerase
or the thermophilic enzymes pfu and vent DNA polymerases.  T4 is con-
venient because it can be heat inactivated prior to ligation.

	
	Let me know if you need more infomation.

	Raj. Odedra
	(mbro at seqnet.dl.ac.uk)




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