Sequencing PCR products after running RT-qPCR is generally unnecessary due to insufficient DNA volume and quantity, which often prevents pipetting the reaction out of the tube. Instead, the correctness of the PCR products can be verified through the design of primers and preliminary tests such as PCR and sequencing. Utilizing RT-PCR products is a viable alternative for further analysis.
PREREQUISITESResearchers in molecular biology, particularly those involved in gene expression analysis and PCR product validation, will benefit from this discussion.