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Nucleic Acid Quantitation with Log-Linear Response Hybridization Probe Sets.

ABSTRACT: 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.

SUBMITTER: Wu LR

PROVIDER: S-EPMC7872156 | biostudies-literature | 2020 Jun

REPOSITORIES: biostudies-literature

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Nucleic Acid Quantitation with Log-Linear Response Hybridization Probe Sets.

Wu Lucia R LR Fang John Z JZ Khodakov Dmitriy D Zhang David Yu DY

ACS sensors 20200612 6

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 ...[more]

PMID: 32475109

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