Why do I have to perform a negative control during SMARTer cDNA synthesis?
Why do I have to perform a negative control during SMARTer cDNA synthesis?
A negative control (performing the entire cDNA synthesis and purification procedure in the absence of any RNA input, but maintaining the same reaction volume) is essential for the evaluation of cDNA synthesis as well as for identifying potential problems, including contamination. We recommend performing a negative control reaction each time the protocol is performed, especially when using the lowest input RNA concentrations.
An example electropherogram of a negative control (no input RNA) using a SMARTer Ultra low kit with 18 cycles of PCR. No contamination is observed.