RT-PCR

[jm]

In article <3oa80j$7f3 at nuscc.nus.sg>, medp4033 at nusunix2.nus.sg (Ramkumar
Lachumanan) wrote:

> Hi netters,
>         I'm having problems with my RT-PCR....think I have internal 
> deletions in the cDNA due to secondary structures during RT-PCR. So, do 
> any of you guys out there have any information on a RT-PCR kit that works 
> at elevated temperatures i.e has a thermostable rev. transcriptase? Am 
> pretty desperate. All help is welcomed!!!!! Thanks.
>                                                     Ram(ses) 
>  I have been using SuperScript II preamplification kit from Life
Technologies(Gibco/BRL). The SuperScript II reverse transcriptase is RNase
H negative so you tend to get longer 1st strand synthesis. I then use
their Taq polymerase (has to be ordered separately). The same buffer is
used for both RT and the PCR.

joe milburn
Dallas VA Med Ctr/UTSWMC