Enzymes activity measurements

[jgasulla]

Hi,

Does someone have protocols to measure the activity of RT or DNA pol?
I saw that NEB’s Unit definition involves the use of radiolabeled dNTPs, like for MMLV:

One unit is defined as the amount of enzyme that will incorporate 1 nmol of dTTP into acid-insoluble material in a total reaction volume of 50 μl in 10 minutes at 37°C using poly(rA) . oligo(dT) as template primer with 50 mM Tris-HCI (pH 8.3), 6 mM MgCl2, 10 mM dithiothreitol, 0.5 mM [3H]-dTTP and 0.4 mM poly(rA) . oligo(dT)12-18. MMLV

Is there any easier way to measure it?

Thanks!
Javier

[osn_scott]

There is a colorimetric enzyme immunoassay for the quantitative determination of retroviral reverse transcriptase activity. It works by incorporation of digoxigenin- and biotin-labeled dUTP into DNA. Here is the bulletin on it from Sigma Aldrich.