T4 RNA Ligase
Dr. Duncan Clark
Duncan at nospam.demon.co.uk
Thu Feb 24 05:37:50 EST 2000
In article <38B4206B.13ACC2C7 at mpiib-berlin.mpg.de>, Marieluise Kirchner
<kirchner at mpiib-berlin.mpg.de> writes
>used anyone the T4 RNA ligase for ssDNA / oligonucleotide-ligation? in the
>buffer of my protocoll hexamine cobalt chloride is used. Why? Is the effect
>really that big? Has hexamine cobalt chloride any bad effects in further
>reactions like PCR or ligation into a vector? The question is - should i use
>this hexamine cobalt chloride or better not?
Have a look at Methods in Enzymology Vol 100. Nothing on the hexamine
but lost on T4 RNA ligase.
On the hexamine cobalt chloride my memory suggests that this was used
many years ago as a ligation promoter for T4 DNA ligase. Having said
that I could be way off!
The problem with being on the cutting edge is that you occasionally get
sliced from time to time....
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