Concentration of DNA in Ligations???

Ned Mantei mantei at
Tue Jul 23 00:37:57 EST 1996

In article <4svv3r$ort at>, M.C.Behrendt at 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   Fax: +41-1-633-1046

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