end point PCR using real time PCR machine (QS5)
Posted 24 June 2020 - 06:15 PM
Our lab only has the QS5 machine for real time PCR. Can this be used to perform endpoint PCR, I would just turn off the camera and unselect sybrgreen is it?
Posted 24 June 2020 - 07:04 PM
Yes, any PCR machine can do end-point PCR... in fact every PCR you do is an end-point PCR because the end-product (DNA band specific for your sequence) is the same whether you are doing qPCR or conventional PCR. In fact, you can even take a qPCR product and run it on a gel to check for single bands. This is how it was done before the days of melt-curves.
There is no need to deselect dyes or anything else, these don't interfere with the production of the DNA.
Posted 26 June 2020 - 12:31 AM
Thank you very much. Also, would it be ok to store the PCR product in the qPCR 96-well plate (sealed and stored in -20deg) for a few days before sending out samples for sequencing.
Posted 26 June 2020 - 02:33 PM
Yes, it should be fine. It's just DNA, so it is quite stable at -20.