Discussion

5 years ago

Multiplex PCR and qPCR

I have been doing multiplex PCR and qPCR separately. Is there a way to combine them in a single reaction using Sybr Green?

Experiment: PCR Multiplex PCR - Poultry DNA

Top comment


That is not possible since multiplex PCR requires different sets of primers and different fluorophores on each matching probe of the sets. Also, you will need to simultaneously detect your different targets via a multichannel qPCR detection system. Using only SYBR green you will only detect it on one specific channel.

Answered 5 years ago


2 Comments


No, it’s not really possible, as multiplex depends on different fluorescent dyes to differentiate between the targets, and Sybr Green fluoresces at just one wavelength, so your instrument will not be able to differentiate between your different targets.

Answered 4 years ago


That is not possible since multiplex PCR requires different sets of primers and different fluorophores on each matching probe of the sets. Also, you will need to simultaneously detect your different targets via a multichannel qPCR detection system. Using only SYBR green you will only detect it on one specific channel.

Answered 5 years ago


Can you help?

Similar discussions


Start your discussion


Share your thoughts or question with experts in your field

Start discussion
Become shareholder Discussions About us Contact Privacy Terms