DNA Ligase in 2nd Strand Buffer. Why?

Hiranya Roychowdhury hroychow at NMSU.EDU
Wed Apr 15 16:10:31 EST 1998

At 11:00 AM 4/15/98 -0700, guggenheim at cf.ac.uk wrote:
>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?

Repairing nicks... If you look up Maniatis you'll see the explanation.
Actually the commercial kits follow the "one tube" RNaseH method that is
described in Sambrook et al. quite elaborately.

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