Concentration of DNA in Ligations???

Ned Mantei mantei at neuro.biol.ethz.ch
Tue Jul 23 00:37:57 EST 1996


In article <4svv3r$ort at bignews.shef.ac.uk>, M.C.Behrendt at sheffield.ac.uk wrote:

> Does anyone know what the optimum concentration of DNA in a ligation 
> reaction is 


See Revie, et al., "Kinetic analysis for optimization of DNA ligase
reactions", Nucleic Acids Res. 16: 10301--10321, 1988. The concentration
you are using may be 10 times too high. My rule of thumb is 1 picomole of
insert and 0.5--10 picomol of vector (the lower amount for large vectors)
per ml, with 10 ul ligations for anything except cDNA banks.

-- 
Ned Mantei
Dept. of Neurobiology, Swiss Federal Institute of Technology
CH-8093 Zurich, Switzerland
mantei at neuro.biol.ethz.ch   Fax: +41-1-633-1046



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