Real Time PCR Probes

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carolabiotech
carolabiotech's picture
Real Time PCR Probes

I would like to know if iyou can use TaqMan probes and MGB probes in the same reaction, different tubes pero the same reaction? Now I am amplifing two exons of a gene designed with MGB probes and I would like to include in the reaction two more exons but the probes for the amplification of those are designed with TAMRA. So, can I amplify all the exons in the same reaction? or I can´t mix the probes? thanks!
 

Ivan Delgado
Ivan Delgado's picture
I am not sure I understand

I am not sure I understand what you are asking. First you state that you want to use both Taqman and MGB probes "in the same reaction, different tubes", which sounds like you want to run each probe separately, in different tubes, and then you ask if you can "amplify all the exons in the same reaction?" and "mix the probes?", which implies you would like to have both probes in the same reaction and the same tube.

Please clarify.

carolabiotech
carolabiotech's picture
 I am sorry, I want to know

 I am sorry, I want to know if I can used for example in one tube the MGB probes and in other tube the TAMRA, and also if I can use them in the same tube....is t clearer now?? :)

Ivan Delgado
Ivan Delgado's picture
It seems you simply want to

It seems you simply want to be able to read MGB and TAMRA probes, either by themselves or in the same tube. In theory you should be able to do this. If you are using an ABI instrument just leave the quencher set to TAMRA. MBG probes do not have a quencher so in theory you can read both signals by assuming the MGB probe will behave just fine using TAMRA settings.

Hope this helps.

carolabiotech
carolabiotech's picture
 Thanks a lot!!!! :)

 Thanks a lot!!!! :)

Ivan Delgado
Ivan Delgado's picture
Anytime. Good luck!

Anytime. Good luck!