DNA Ligase in 2nd Strand Buffer. Why?

Dr. Duncan Clark duncan at genesys.demon.co.uk
Thu Apr 16 03:41:27 EST 1998

In article <55FC532AA at RNCHCF2S.CF.AC.UK>, Jeremy A. Guggenheim
<Guggenheim at cardiff.ac.uk> writes
>Several manufacturers (e.g. Gibco BRL) use E. Coli DNA Ligase in their
>second strand cDNA reaction buffers (along with the usual DNA
>ploymerase and RNase H). They then say that the products formed will
>be blunt-ended, double stranded cDNAs.
>Does anyone know what the DNA ligase is there for?
>And couldn't it cause concatemerization of the products?

Sealing nicks. Unlike T4 DNA ligase, E.coli DNA ligase appears to be
much more limiting in what overhangs etc it will ligate.

The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
DNAmp Ltd.
TEl/FAX 01252376288

More information about the Methods mailing list