Vol of ligation mix to use in transformation

David Micklem seemysignature at mole.bio.cam.ac.-uk
Thu Jan 30 07:23:01 EST 1997


In article <32F05C68.4369 at sheffield.ac.uk>, K.Mulcahy at sheffield.ac.uk wrote:


>Also, does anyone know the answer to the following question? When adding 
>your ligation mix to the bacteria (without precipitation of the DNA 
>first) why is it recommended (e.g. in the Gibco T4 DNA ligase protocol) 
>that you dilute the ligation mix more than 5 times before adding it to 
>the bacteria?
>
>Many thanks,
>
>Kevin Mulcahy.

Hi Kevin,

There was a BRL Focus article (ie Gibco!) on optimizing ligation
conditions. They showed that diluting the ligation mix results in a much
higher transformation efficiency.  (Presumably there is some inhibitory
component in ligation mix).

I can probably dig out the exact reference if you want it.

David

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