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A single nucleotide polymorphism or SNP is a single nucleotide variation at a specific genomic position in a large population. It is the most prevalent type of sequence variation found in the human genome. Point mutations that occur in more than 1% of the population qualify as SNPs. These are present once every 1000 nucleotides on an average in the human genome. Replacement of a purine with another purine (A/G) or a pyrimidine with another pyrimidine (C/T) is known as a transition. In contrast,...
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Multiplex Single Nucleotide Polymorphism Analyses.

Steve F C Hawkins1, Paul C Guest2

  • 1Bioline Reagents Limited, Unit 16, The Edge Business Centre, Humber Road, London, NW2 6EW, UK. shawkins@bioline.com.

Methods in Molecular Biology (Clifton, N.J.)
|November 30, 2016
PubMed
Summary
This summary is machine-generated.

This study introduces a new method for simultaneous gene expression analysis using quantitative polymerase chain reaction (qPCR) with up to five fluorescent probes. This technique enhances real-time detection and quantitation of multiple amplicons efficiently.

Keywords:
AlleleFluorescent dyesMultiplex analysisPCRQuantitationSingle nucleotide polymorphismTaq polymerase

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Quantitative polymerase chain reaction (qPCR) is a standard technique for real-time gene expression analysis.
  • Fluorescent reporter probes are incorporated into amplified cDNA strands, with signal intensity correlating to reaction progress.
  • Multiplexing, using probes with distinct fluorescent wavelengths, enables simultaneous detection of multiple targets.

Purpose of the Study:

  • To present a novel method for simultaneous real-time quantitation of multiple gene targets.
  • To demonstrate the utility of SensiFAST and SensiFAST One-Step probe kits for multiplex qPCR.

Main Methods:

  • Utilized SensiFAST and SensiFAST One-Step probe kits for quantitative polymerase chain reaction.
  • Employed fluorescent reporter probes with distinct wavelengths for multiplexing.
  • Performed simultaneous real-time quantitation of up to 5 amplicons.

Main Results:

  • Successfully demonstrated simultaneous real-time quantitation of up to 5 amplicons.
  • Validated the effectiveness of the SensiFAST and SensiFAST One-Step probe kits for multiplex qPCR applications.

Conclusions:

  • The described method enables efficient and simultaneous real-time quantitation of multiple gene targets.
  • SensiFAST and SensiFAST One-Step probe kits provide a robust solution for multiplex qPCR assays.