Engineering Commons^™

Novel Kinetic Description Of Real-Time Polymerase Chain Reaction Characterizes Interrelated Effects Of Sample, Master Mix, And Cycle Time, David Tafur

All Graduate Theses and Dissertations, Fall 2023 to Present

Real-time quantitative polymerase chain reaction fluorescent curves are influenced by the quantification method, template length, primer sequence, polymerase activity, reaction conditions, and unwanted side reactions. Interpretation of these curves depends on understanding the subjacent mechanisms by which those factors can alter the curve profile, particularly on highly diluted analytes or impure samples. However, such factors are individually described today, or their interplay is not fully developed. The present work examines the effect of primer dimer formation and extension, resource competition, template reannealing, and polymerase thermodegradation, in addition to novel descriptions of polymerase competitive inhibition, extended primer amplification, and the probability …