pre-incubation at 30 for RTs using random hexamers

Alexander Dobrovic alexander.dobrovic at adelaide.edu.au
Mon Sep 4 21:26:56 EST 2000


Roche recommend a 10 minute pre-incubation at 30C prior to a 45 minute
incubation at 42C if random haxamers are used with their EXPAND Reverse
transcriptase. Presumably, this is due to the very low Tms of random
hexamers. Is this a balance between the Tms and the transcriptase activity?
This follows a 65C heating for 10 minutes - will the RNA renature after
this?

I would be grateful for people's opinions as to whether this does in fact
make a difference. We'll also test this out and respond back to the group.

Alexander Dobrovic, Ph.D.
Chief Medical Scientist/Dept of Haematology-Oncology
Affiliate Senior Lecturer/University of Adelaide Dept of Medicine
The Queen Elizabeth Hospital
Woodville, SA 5011, Australia

Tel  61-8-8222 7676
Fax  61-8-8222 6046
alexander.dobrovic at adelaide.edu.au



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