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Related Concept Videos

Single Nucleotide Polymorphisms-SNPs01:05

Single Nucleotide Polymorphisms-SNPs

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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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Related Experiment Video

Updated: Aug 10, 2025

Infinium Assay for Large-scale SNP Genotyping Applications
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Published on: November 19, 2013

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Amplifluor-Based SNP Genotyping.

Manmode Darpan Mohanrao1, Senapathy Senthilvel1, Yarabapani Rushwanth Reddy1

  • 1ICAR-Indian Institute of Oilseeds Research, Hyderabad, India.

Methods in Molecular Biology (Clifton, N.J.)
|February 13, 2023
PubMed
Summary

Amplifluor is a genotyping system for analyzing single nucleotide polymorphisms (SNPs) using polymerase chain reaction (PCR). This method employs allele-specific primers and fluorescence detection for SNP genotyping.

Keywords:
AmplifluorGenotypic assaysSingle nucleotide polymorphisms (SNPs)

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Single nucleotide polymorphisms (SNPs) are key genetic variations.
  • Accurate SNP genotyping is crucial for various biological and medical studies.
  • Existing methods may require specific equipment or complex protocols.

Purpose of the Study:

  • To present the principle and working protocol of the Amplifluor assay.
  • To demonstrate SNP allele analysis using end-point fluorescence detection.
  • To provide a practical example of the Amplifluor assay execution.

Main Methods:

  • The Amplifluor assay utilizes polymerase chain reaction (PCR).
  • It employs two competing allele-specific primers and a common reverse primer.
  • Analysis involves end-point fluorescence detection using a plate reader or real-time measurement with a qPCR machine.

Main Results:

  • The Amplifluor system enables the analysis of single nucleotide polymorphisms (SNPs).
  • The assay relies on allele-specific primer extension and subsequent fluorescence detection.
  • Successful genotyping of SNPs is achievable with sequence information and appropriate detection equipment.

Conclusions:

  • The Amplifluor assay provides a robust method for SNP genotyping.
  • The protocol is adaptable for end-point fluorescence detection.
  • This system offers a valuable tool for genetic analysis and research.