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end point PCR using real time PCR machine (QS5)


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#1 SF_HK

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Posted 24 June 2020 - 06:15 PM

Hi, I need to perform end Point PCR on my targets to send them out for sequencing.

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?

many thanks

#2 bob1

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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.



#3 SF_HK

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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.



#4 bob1

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Posted 26 June 2020 - 02:33 PM

Yes, it should be fine. It's just DNA, so it is quite stable at -20.






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