What concentrations of reagents are recommended to start standardizing an RT-PCR primer set?
To start standardizing an RT-PCR primer set, it's recommended to test a range of primer concentrations, typically between 100 nM and 900 nM. This allows you to find the optimal concentration that produces efficient and specific amplification of the target gene.
To begin standardizing an RT-PCR primer set, start with primer concentrations ranging from 100 nM to 900 nM. This will help determine the best concentration for efficient and specific target gene amplification.
The only components you can vary are the amount of cDNA and primer concentration.
For everything else, follow the instructions in the kit since those are already optimized (e.g. temps to use for polymerase).
Amount of cDNA is going to depend on expression of your gene-of-interest so you'll have to figure it out by trial and error.
Good luck!
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