Fast Optimization of a Multiplex Influenza Identification Panel Using a Thermal Gradient

This tech report focuses on the use of the thermal gradient feature on the CFX96™ real-time PCR detection system to optimize reverse transcription (RT) and fast PCR conditions for a four-target multiplex PCR assay. It is demonstrated that the gradient feature of the CFX96 system dramatically accelerates the optimization of the RT and PCR steps for multiplexing RT-qPCR assays and improves the ease of converting cycling conditions from a three-step to a two-step PCR assay.