dithiothreitol for RT

Nick Theodorakis nick_theodorakis at hotmail.com
Wed Dec 15 21:44:51 EST 2004


On Wed, 15 Dec 2004 22:49:43 +0100, fabio <fabio at fabius.it> wrote:

>I have faund the following mix for RT:
>RNA tamplate, random hexameres, --> 70°C x 10'-->ice ,first-strand 
>buffer (Invitrogen), dithiothreitol, dNTPs, RNAsin, M-MLV-RT -->37°C x 
>1h-->70 °C x 10 ' --> PCR.
>The question is: why dithiothreitol ?

Among other things, RNasin requires DTT to remain active. I don't know
if RT requires it, but in general, many, if not most, intracellular
enzymes prefer to be in a little reducing environment.

Nick

-- 
Nick Theodorakis
nick_theodorakis at hotmail.com
contact form:
http://theodorakis.net/contact.html



More information about the Methods mailing list