The kinetics of polymerase chain reaction product duplex dissociation

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Title The kinetics of polymerase chain reaction product duplex dissociation
Publication Type thesis
School or College College of Engineering
Department Biomedical Engineering
Author Betz, Daniel
Date 2016
Description The overall speed of a polymerase chain reaction (PCR) is determined by many factors including the time required for denaturation during each cycle. Indirect evidence suggests that denaturation during PCR is complete in less than a second. Therefore, our experimental observations of DNA denaturation were conducted with a stop-flow instrument having a ultrashort time resolution of 1.2 ms. Duplex denaturation under PCR conditions was monitored with a fluorescent dye using PCR products of 51, 100, 272, and 547 bp. The 100 bp product showed a t1/2 of 100 ms when the temperature was 2.4 C above the Tm. Rates increased rapidly at higher temperatures. However, rates did not correlate with the length of the PCR product; the longest (547 bp) product had 2 melting domains. Our results suggest that adequate denaturation time and temperature control are necessary refinements for successful high-speed PCR.
Type Text
Publisher University of Utah
Subject Pure sciences; Applied sciences
Dissertation Name Master of Science
Language eng
Rights Management ¬©Daniel Betz
Format application/pdf
Format Medium application/pdf
ARK ark:/87278/s6tx7kbf
Setname ir_etd
ID 1343459
Reference URL https://collections.lib.utah.edu/ark:/87278/s6tx7kbf
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