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Related Concept Videos

PCR01:32

PCR

Overview
RACE - Rapid Amplification of cDNA Ends02:35

RACE - Rapid Amplification of cDNA Ends

Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific primer.
Since the...

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Related Experiment Video

Updated: Jun 20, 2026

Primer Extension Capture: Targeted Sequence Retrieval from Heavily Degraded DNA Sources
15:28

Primer Extension Capture: Targeted Sequence Retrieval from Heavily Degraded DNA Sources

Published on: September 3, 2009

Modified multiple primer extension method.

Toshihide Yanagawa1, Hisashi Koga

  • 1Laboratory of Medical Genomics, Department of Human Genome Technology, Kazusa DNA Research Institute, Kisarazu, Chiba, Japan.

Methods in Molecular Biology (Clifton, N.J.)
|September 22, 2009
PubMed
Summary
This summary is machine-generated.

This study refines the multiple primer extension (MPEX) method for enhanced single nucleotide polymorphism (SNP) detection. The modified MPEX method utilizes a novel substrate and visualization technique for accurate genetic analysis.

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Last Updated: Jun 20, 2026

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15:28

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Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis
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Published on: June 19, 2018

Candidate Gene Testing in Clinical Cohort Studies with Multiplexed Genotyping and Mass Spectrometry
05:53

Candidate Gene Testing in Clinical Cohort Studies with Multiplexed Genotyping and Mass Spectrometry

Published on: June 21, 2018

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Multiple primer extension (MPEX) is a technique for DNA template amplification.
  • Existing MPEX methods have limitations in sensitivity and specificity for certain applications.
  • Single nucleotide polymorphisms (SNPs) are crucial genetic markers for various traits and diseases.

Purpose of the Study:

  • To modify and optimize the MPEX method for sensitive and specific single nucleotide polymorphism (SNP) detection.
  • To develop a robust platform for allele-specific SNP analysis.
  • To demonstrate the utility of the modified MPEX method in analyzing clinically relevant genes.

Main Methods:

  • A modified multiple primer extension (MPEX) method was developed using a plastic S-BIO PrimeSurface substrate.
  • Hybridization and extension reactions were optimized on the specialized substrate.
  • Biotin-dUTP incorporation and a colorimetric detection system (BCIP/NBT) were employed for visualization.
  • The method was combined with multiplex PCR for simultaneous analysis of multiple SNPs.
  • The modified MPEX method was applied to analyze single nucleotide polymorphisms (SNPs) in the micro opioid receptor gene (OPRM1).

Main Results:

  • The modified MPEX method demonstrated stable thermal and chemical properties on the PrimeSurface substrate.
  • Allele-specific PCR products were successfully visualized using biotin-dUTP and colorimetric detection.
  • The combined multiplex PCR and modified MPEX method enabled efficient analysis of multiple SNPs.
  • Representative SNPs within linkage disequilibrium blocks of the OPRM1 gene were accurately analyzed.
  • The method showed high specificity and sensitivity for single nucleotide polymorphism (SNP) detection.

Conclusions:

  • The modified MPEX method offers a refined approach for single nucleotide polymorphism (SNP) detection with improved performance.
  • The use of the S-BIO PrimeSurface substrate and colorimetric detection enhances the reliability and ease of SNP analysis.
  • This optimized method, particularly when combined with multiplex PCR, provides a powerful tool for genetic research and diagnostics, exemplified by its application to the OPRM1 gene.