Concentration of DNA in Ligations???

Paul N Hengen pnh at cockleberry.ncifcrf.gov
Tue Jul 23 12:29:20 EST 1996


Ned Mantei (mantei at neuro.biol.ethz.ch) 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.

One of the best introductory explanations I found on this subject is in the
book by Rodriguez, R. L., and R. C. Tait. 1983. Recombinant DNA techniques:
An introduction. Benjamin/Cummings Publishing Co., Inc. Menlo Park, CA.

--
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