Ask for help on RT-PCR
yww at CIMS1.YZU.EDU.CN
Fri Mar 6 09:04:13 EST 1998
Last September I succeeded in amplifying a 1.8 Kb long viral RNA by RT-PCR=
using 1st strand cDNA synthesis kit for RT-PCR and PCR Master (both made=
by Boehringer Mannheim ) with the PCR machine( Perkin-Elmer) resulting in=
the only 1.8 Kb product. But this month I can=92t repeat the result , that=
is to say I got nothing . The only difference between these two runs is=
the RNA sample which isolate by Promega=92s Rnagents Total RNA Isolation=
System at different time (Just before RT-PCR each time). Can someone give=
me some good advises on how to repeat the RT-PCR ?
And now I still have some product got from last RT-PCR . Can I reamplify=
the product and how to do ?
The last question is how long can I store the viral RNA isolated by=
Promega=92s Rnagents Total RNA Isolation System under =9630 degree C in=
Rnase freee water. (I want to reuse the RNA sample got in last September,=
because it is now very hard to get the virus. )
Thanks in forward and waiting for your good advises eagerly .
Yww at cims1.yzu.edu.cn
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