PCR Multiplexing Based on a Single Fluorescent Channel Using Dynamic Melting Curve Analysis
Haoqing Zhang, Martina Gaňová, Zhiqiang Yan, Honglong Chang, Pavel Neužil
Abstract
and then used the data to construct individual PCR amplification curves for each gene to determine the initial concentration of DNA in the sample. Our proposed method allows researchers to look inside the PCR in each thermal cycle, determining the PCR product specificity in real time instead of waiting until the end of the PCR. Additionally, the slow transition rate from elongation to denaturation provides a dynamic multiplexing assay, allowing the detection of at least three genes in real time.
Topics & Concepts
Melting curve analysisSYBR Green IPrimer dimerReal-time polymerase chain reactionTaqManMolecular biologyDNAPolymerase chain reactionFluorescenceChemistryBiologyGeneBiochemistryMultiplex polymerase chain reactionQuantum mechanicsPhysicsMolecular Biology Techniques and ApplicationsBiosensors and Analytical DetectionAdvanced Biosensing Techniques and Applications