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Related Concept Videos

Sanger Sequencing01:57

Sanger Sequencing

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DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
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Next-generation Sequencing03:00

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The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....
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Targeted Amplicon Genotyping by Sanger Sequencing.

Anthony Torres1, Reginald Gaudino2,1

  • 1Cannabis Research Institute, Discovery Partners Institute, University of Illinois System, Chicago, IL, USA.

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|June 28, 2025
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Summary

Sanger sequencing is a cost-effective, accurate method for targeted DNA sequencing. This protocol details primer design, chain termination, electrophoresis, and chromatogram analysis for identifying genetic variations.

Keywords:
AmpliconChromatogram analysisDNAElectrophoresisEnd termination reactionSNP detectionSanger sequencingSingle nucleotide polymorphism (SNP)Targeted sequencing

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Sanger sequencing remains a valuable tool for targeted DNA analysis.
  • Conventional methods offer a balance of speed, cost-effectiveness, and accuracy.

Purpose of the Study:

  • To present a detailed protocol for performing Sanger sequencing.
  • To outline the steps for accurate base identification and variation detection.

Main Methods:

  • Designing specific primers for DNA amplification.
  • Incorporating fluorophore-labeled, chain-terminating dideoxynucleotides.
  • Separating DNA fragments via electrophoresis and analyzing chromatograms.

Main Results:

  • Precise base identification within DNA amplicons.
  • Detection of nucleotide changes indicative of genetic variations.
  • Facilitation of amino acid variation analysis in targeted genes.

Conclusions:

  • Sanger sequencing provides a reliable method for targeted genetic analysis.
  • The protocol enables accurate identification of nucleotide and amino acid variations.
  • This technique remains relevant for cost-effective and rapid genetic studies.