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?
>
>Jez
>
>

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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