Nucleic Acid Quantitation with Log-Linear Response Hybridization Probe Sets
Concentrations of different nucleic acid species in biological samples span many orders of magnitude. A real-time polymerase chain reaction maps the concentration of a target nucleic acid sequence log-linearly into cycle threshold to enable quantitation with a wide dynamic range but suffers from enzymatic biases. Here, we present a general design for constructing hybridization probe sets with highly log-linear response curves to enable accurate enzyme-free quantitation across large ranges (more than 6 logs) of target DNA concentrations. The sensitivity of each component probe is accurately adjusted via formulation stoichiometry to reduce the standard error of target quantitation down to 7%. As a proof of concept, we show multiplexed quantitation of three microRNA species in total RNA of the human brain and liver.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2020 |
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| Erschienen: |
2020 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:5 |
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| Enthalten in: |
ACS sensors - 5(2020), 6 vom: 26. Juni, Seite 1604-1614 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Wu, Lucia R [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 14.05.2021 Date Revised 14.05.2021 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1021/acssensors.0c00052 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM310555515 |
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| 520 | |a Concentrations of different nucleic acid species in biological samples span many orders of magnitude. A real-time polymerase chain reaction maps the concentration of a target nucleic acid sequence log-linearly into cycle threshold to enable quantitation with a wide dynamic range but suffers from enzymatic biases. Here, we present a general design for constructing hybridization probe sets with highly log-linear response curves to enable accurate enzyme-free quantitation across large ranges (more than 6 logs) of target DNA concentrations. The sensitivity of each component probe is accurately adjusted via formulation stoichiometry to reduce the standard error of target quantitation down to 7%. As a proof of concept, we show multiplexed quantitation of three microRNA species in total RNA of the human brain and liver | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a RNA quantitation | |
| 650 | 4 | |a dynamic range | |
| 650 | 4 | |a enzyme-free biosensor | |
| 650 | 4 | |a nucleic acid hybridization | |
| 650 | 4 | |a nucleic acids thermodynamics | |
| 650 | 4 | |a toehold probe | |
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| 700 | 1 | |a Fang, John Z |e verfasserin |4 aut | |
| 700 | 1 | |a Khodakov, Dmitriy |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, David Yu |e verfasserin |4 aut | |
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