Hello!
I am trying to find more resources on how to perform Multiplex Analyses Using Real-Time Quantitative PCR on brain tissue from punches (.5mm). Because the tissue is so small, the cDNA yield is very low therefore I am only able to check for 1 or 2 genes being expressed per brain tissue sample. I would like to check for the differential expression of more genes running a multiplex analyses using RT-qPRC with the LightCycler 480 system.
Please let me know if you have more information/protocols on this system.
Thank you in advance
Hi Vanessa Alizo , I perform the multiplex qPCR on functional genes involved on the nitrogen cycles. When you find the correct primer to use (that is possible to use together). When you find, you need to design the probe and after to test the 1 gene alone and together the 2 gene to optimase the qPCR (in this case the 1 gene must have the same CT in both the conditions).
Hi Giorgia Pertile thank you for the reply. Would that mean both genes will be using the same primer, but different probes ? Also, are you able to differentiate between the 2 genes?
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