efficient blunt ligation?

Mike Holloway mike.holloway at stjude.org
Mon Jun 26 13:33:31 EST 1995


In article <3sdpek$n42 at worak.kaist.ac.kr>, hbkim at biochem.kaist.ac.kr (Hanbok Kim) says:
>
>Is there any effective method for blunt end ligation
>by temperature regulation 

Try the Boehringer Mannheim Rapid DNA ligation kit.  Their buffer comes 
with some viscous secret ingredient (PEG + something else?) and a fairly 
high concentration ligase (5u/ul).  Instructions say 5 minutes at room temp,
sticky or blunt end.  So far, its working as advertised.  They also insist
that you use high efficiency hexamminecobalt chloride method competent 
cells, which would have an affect on the success of any ligation project.

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