So my lab recently switch from TaqMan Gene Expression Assay Mix (20X) to using custom probes and primers from IDT. That being said the primers and probes all came separately instead of as a mix. It also did not come with any protocols and I can't seem to find any on IDTs website.
So the TaqMan Assay Mix states that it has the following concentrations: 18uM Forward Primers, 18uM Reverse Primers, and 5uM Probe. The custom primers and probes from IDT came in the same concentrations: 18uM Forward Primers, 18uM Reverse Primers, and 5uM Probe.
So to make my 20X Assay Mix with my custom reagents do I just combine my primers and probe in equal parts? So for example, I would mix 1uL of forward primer, 1uL of reverse primer, and 1uL of probe then use as normal?
In my case with the Taqman work if I prepare separately the probe and the primer. I prepare the primer at 20 uM and the probe at 10 uM and after I make thae calculation and I add the volume for each thing to obtain a final concentration of 0.5 uM for the probe and 0.5 uM for the primer inside the mix. Amanda Harmon
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