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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

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Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
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DNA Microarrays02:34

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Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
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In-situ Hybridization02:31

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In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
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Labeling DNA Probes03:31

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DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
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Related Experiment Video

Updated: Dec 20, 2025

Split Hybridization Probe Utilizing a DNA Fluorescent Light-up Aptamer as a Signal Reporter for Sequence-Specific Nucleic Acid Analysis
07:10

Split Hybridization Probe Utilizing a DNA Fluorescent Light-up Aptamer as a Signal Reporter for Sequence-Specific Nucleic Acid Analysis

Published on: July 8, 2025

903

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

Lucia R Wu1, John Z Fang1, Dmitriy Khodakov1

  • 1Department of Bioengineering, Rice University, Houston, Texas 77005, United States.

ACS Sensors
|June 2, 2020
PubMed
Summary
This summary is machine-generated.

This study introduces enzyme-free DNA quantitation using novel hybridization probes. These probes offer accurate measurements across a wide concentration range, improving upon traditional methods.

Keywords:
RNA quantitationdynamic rangeenzyme-free biosensornucleic acid hybridizationnucleic acids thermodynamicstoehold probe

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genomics

Background:

  • Nucleic acid concentrations vary greatly in biological samples.
  • Real-time polymerase chain reaction (PCR) offers wide dynamic range quantitation but has enzymatic biases.

Purpose of the Study:

  • To develop a general design for hybridization probe sets enabling accurate, enzyme-free quantitation of target DNA.
  • To achieve highly log-linear response curves for precise measurements across extensive concentration ranges.

Main Methods:

  • Designed hybridization probe sets with sensitivity adjusted via formulation stoichiometry.
  • Demonstrated multiplexed quantitation of microRNA species in human brain and liver total RNA.

Main Results:

  • Achieved highly log-linear response curves for probe sets.
  • Enabled accurate enzyme-free quantitation across more than 6 logs of target DNA concentrations.
  • Reduced the standard error of target quantitation to as low as 7%.

Conclusions:

  • The developed probe design facilitates accurate, enzyme-free DNA quantitation.
  • This method overcomes limitations of enzymatic biases found in real-time PCR.
  • Successfully demonstrated multiplexed microRNA quantitation in human tissues.