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231225s2020 xx |||||o 00| ||eng c |
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|a 10.2144/btn-2020-0016
|2 doi
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|a eng
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1 |
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|a Warton, Kristina
|e verfasserin
|4 aut
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|a Target sequence heterogeneity causes the 'hook effect' in fluorescent dye-based quantitative PCR
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|c 2020
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|a Text
|b txt
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|a ƒaComputermedien
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|a ƒa Online-Ressource
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|a Date Completed 21.06.2021
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|a Date Revised 21.06.2021
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|a published: Print-Electronic
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|a Citation Status MEDLINE
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|a The 'hook effect' describes a phenomenon in quantitative PCR (qPCR) amplification curves where fluorescence values decrease following an initial amplification phase. We propose that in intercalating dye-based qPCR, the 'hook effect' is due to the amplification of heterogeneous but related DNA targets. The decrease in fluorescence at later cycles occurs because the related products self-anneal to form a DNA heteroduplex with a melt temperature below the temperature at which the fluorescence measurement is made. We show this experimentally using qPCR of Alu family repetitive DNA elements
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|a Letter
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|a Research Support, Non-U.S. Gov't
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|a hook effect
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|a optimization
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|a quantitation
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|a quantitation artifact
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|a quantitative PCR
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|a Fluorescent Dyes
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|a Intercalating Agents
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|a DNA
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1 |
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|a Xu, Yue
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|a Ford, Caroline Elizabeth
|e verfasserin
|4 aut
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| 773 |
0 |
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|i Enthalten in
|t BioTechniques
|d 1988
|g 69(2020), 2 vom: 07. Aug., Seite 80-83
|w (DE-627)NLM012627046
|x 1940-9818
|7 nnns
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| 773 |
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|g volume:69
|g year:2020
|g number:2
|g day:07
|g month:08
|g pages:80-83
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|u http://dx.doi.org/10.2144/btn-2020-0016
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