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why need the "Data collection step" - (May/01/2007 )

I got a rt-PCR protocol which includes a 79C 27 sec step for data collection. Could anyone told me why this step is necessary? Thanks!

-mindy-

QUOTE (mindy @ May 2 2007, 05:52 AM)
I got a rt-PCR protocol which includes a 79C 27 sec step for data collection. Could anyone told me why this step is necessary? Thanks!


My guess it is from a real time PCR protocol. If you are not doing real time PCR then you should be able to avoid using it.

Cheers

Daniel

Longer ABI sequencing traces

-Daniel Tillett-

Thank!!
Yes. I am doing rt-PCR. But my understanding is that even rt-PCR does not always have this step. Do you know in which situation you need to have this step?

-mindy-

Are you sure that it is a data collection step? Sounds like an extension step and the data collection is at the end. If you are doing rt-PCR (=reverse transcription), you do not need to collect data. If you use real-time PCR (or even real-time RT-PCR) you definetly want to collect the data after each cycle - otherwise real-time makes no sense. There are protocols using a combined annealing/extension step (so called two step protocol). You need to provide more information if you want to know if your PCR is suitable for a two-step approach.

BEst wishes for your amplification, Kr├╝mel

-kr├╝melmonster-