RT-PCR troubleshooting

Sarah Roberts-Thomson s.roberts-thomson at PHARMACY.UQ.EDU.AU
Tue Jun 1 17:37:07 EST 1999


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Hi,

I am trying to amplify a particular mRNA target and having little
success, although other targets cause us no problems. I know the mRNA is
present in the sample RNA we are using. We are using a one step RT-PCR
kit and can show that the PCR step is working with our primers by using
a brain cDNA library. We also know that all the components in this kit
are okay as we can use it to amplify other targets. Does anyone have any
suggestions? We have tried a range of magnesium conc, annealing temp etc
etc.

Thanks

Sarah


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