Insert size?

James S. Sutcliffe jamess at mbcr.bcm.tmc.edu
Tue Nov 23 09:10:15 EST 1993


Yet another followup to this insert size issue:

Tony was quite correct in his statement about the correct molar ratios of
the fragment ends. Another thing that might help would be to do a two step
ligation, where the first "step" is a short (e.g. 2-3 h) at a higher
concentration to promote intermolecular ligation events and the second
"step" is at a more dilute concentration to promote intramolecular ligation
events. Don't forget that transformation efficiency is inversely
proportional to the size of the construct. Hope this helps.

Jim Sutcliffe
Institute for Molecular Genetics
Baylor College of Medicine



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