Any experience in improving blunt end ligation?

Don Walthers genejcky.removethis at yahoo.com
Fri Jun 29 18:31:05 EST 2001


Try lowering the ATP concentration 10 fold. For NEB ligase, I just dilute
the 10X ligase buffer with NEB#2 10X (or whatever is closest, although
ligase works in all RE buffers). Also do the ligation at RT as opposed to 16
degrees. Rouche also has a 5X concentrated ligase.

--
Don Walthers, PhD genejcky @ yahoo com
Dept of Molecular Microbiology
Oregon Health Sciences University
Portland OR
Our gene pool needs a little chlorine.





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