How do I know if my first-cycle cDNA synthesis reaction has worked well?
It is challenging to assess the success of the cDNA synthesis reaction and this applies to both the One-Cycle and Two-Cycle protocol. This is because the amount of double-stranded cDNA obtained is quite small, and it is mixed in with the starting total RNA materials. The best way to QC this step is to carry out the subsequent IVT reaction and quantitate the cRNA yield.