We are running a MMQPCR on bovine telomeres and have 2 absolute standards, one for telomeres, and one for our single-copy gene. We've confirmed both primers and standards work and give good R^2 and slopes, but we are wondering about the accuracy and efficiency of having both standards in the same wells with our primer cocktail. Has anyone combined the standards and measured for 2 Std. curves? This is something that is usually too detailed for publications.
Hi
In case you mean multiplex PCR I can say that this should be an issue as long as the mastermix is capable of multiplexing. Hope I understood your question well let me know if you need more details
Sven
Unless your different products are somehow labeled differently, there will be no way to differentiate the two different products. I've never seen a publication with multiplexed qPCR samples, but if you can find one, please share.
In my experience, you should do a multiplex validation study. Perform a run with your samples and standards in singleplex while (in parallel) running the same samples but in multiplex, and see whether or not anything has changed.
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