AMV vs. MMLV reverse transcriptase

Neil Saunders saunders at bio.vu.nl
Wed Nov 18 09:52:15 EST 1998



Marc Gregory Busch wrote:

>     I am currently trying to made a cDNA library of total RNA using random
> hexamer primers. The old protocol uses MMLV. However AMV reverse
> transcriptase is cheaper. Is there a difference and what's better for
> certain conditions. After looking the most I could find is that AMV is
> stable at a slightly higher temperature. Thank you an advance for any
> assistance you may have in the manner.
>                                                                     Marc
> Gregory Busch
>
> mgbusch at ucdavis.edu

As I remember, AMV-RT is more processive than MMLV-RT, meaning that less units
are required in the reaction; I think typically 1 microlitre of MMLV-RT in a
reaction is about 200U, whereas 1 microlitre of AMV-RT is a few tens of units
(depending on the concentration from the supplier, obviously).  AMV-RT is
supposed to be more stable at higher temperatures, meaning that you should be
able to raise the incubation temperature and so lessen the truncation of
cDNA's due to pausing at secondary structure, although MMLV-RT is also usable
to at least 42C.  I've used them both, for 5'-RACE, primer extension and
RT-PCR and have found one about as good as the other, to be honest.

Neil Saunders

--
Department of Molecular Cell Physiology,
Faculty of Biology,
Vrije Universiteit,
De Boelelaan 1087,
1081 HV, Amsterdam
The Netherlands

phone: +31 20 4447194
email: saunders at bio.vu.nl
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