blunt-end ligation

Roland Hübner rhubner at gins.uia.ac.be
Fri Oct 27 04:30:57 EST 2000


Hello,

> T4 ligase protocol has the following modifications

 not much difference here IMHO

> > 5ng vector (dephosphorylated)
> > 5ng insert
> 
> 3-10 x more insert than vector

 insert is often 3-10 x shorter...

> > 2uL of the 5x ligation buffer
> > 1 U ligase
> 
> 4U

 stock concentrations? unit definitions?
 
> > total vol. to 10uL
> 
> add 1/10 vol 50% PEG4000 (endconc. 5%)

 PEG is included in the buffer described in Focus...

Cheers,
Roland






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