RT with biotin dNTP's - Reply

Varyc varyc.mmcri at OFFICE.MMC.ORG
Wed Feb 1 06:59:31 EST 1995


Hi all,
Z-Suldan writes:

>How about the RT enzyme?? We normally use BRL's Superscript
>here for
>normal cDNA synthesis, but maybe there's a better one when
>biotin dUTP
>is in the reaction??

I was under the impression that RT doesn't like messed up
dNTP's. The RT proofreading mechanism (what there is of one)
relies on kinetic barriers (i.e. high off rate) resulting in no
significant binding of "wrong" dNTPs; especially bio- or other
substituted dNTPs. 

T.aq., calf pol alpha and othe DNA pols w/o 3'exo activity will,
however, take bio-dNTPs. I'm not sure about Tth polymerase but
since it has no 3'exo it may be picky as well.

Be interested to know what people think on this,

Cal Vary



More information about the Methods mailing list