Dear all, I need to run a semi-quantitative style PCR, not for quantification, but just to monitor the progress of a multiplex reaction. When PCR tubes are removed at the end of a particular cycle, what is the most common way to cool them down and stop the reaction progress? Is transferring to a PCR block at 4 degrees enough, or do I need ice, or even dry ice? Many thanks in advance.
Laurence Stuart Dawkins-Hall
Hi Martha. I would:
- Pause the PCR block and remove tube
- transfer onto ice and whilst coooling add 1/10 volume TE buffer
- Transfer to -20C freezer until such time as you are ready to analyse
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Igor Kurochkin
I usually put it to store to a refrigerator (4oC). The reaction would not proceed anyway as long as DNA polymerase does not work. And it does not work at 4oC.
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