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Sanger Sequencing

Sanger Sequencing

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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Related Experiment Video

Updated: Jul 29, 2025

Detecting Somatic Genetic Alterations in Tumor Specimens by Exon Capture and Massively Parallel Sequencing

Detecting Somatic Genetic Alterations in Tumor Specimens by Exon Capture and Massively Parallel Sequencing

Published on: October 18, 2013

Nucleotide-selective amplification and array-based detection for identifying multiple somatic mutations.

Luis A Tortajada-Genaro¹, Ana Lazaro², Sara Martorell²

¹Instituto Interuniversitario de Investigación de Reconocimiento Molecular y Desarrollo Tecnológico (IDM), Universitat Politècnica de València, Universitat de València, Camino de Vera s/n, 46022, Valencia, Spain; Unidad Mixta UPV-La Fe, Nanomedicine and Sensors, Valencia, Spain; Departamento de Química, Universitat Politècnica de València, Valencia, Spain.

Analytica Chimica Acta

|May 25, 2023

Summary

This summary is machine-generated.

A new genotyping tool detects single-nucleotide changes for personalized cancer therapy. This cost-effective biosensing method accurately identifies mutations in genes like KRAS from patient samples.

Keywords:

Allele-specific techniques Array sensing DNA chip technology Mutations in oncogenes

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Oncogenic Gene Fusion Detection Using Anchored Multiplex Polymerase Chain Reaction Followed by Next Generation Sequencing

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Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations

Published on: March 29, 2017

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Last Updated: Jul 29, 2025

Detecting Somatic Genetic Alterations in Tumor Specimens by Exon Capture and Massively Parallel Sequencing

Detecting Somatic Genetic Alterations in Tumor Specimens by Exon Capture and Massively Parallel Sequencing

Published on: October 18, 2013

Oncogenic Gene Fusion Detection Using Anchored Multiplex Polymerase Chain Reaction Followed by Next Generation Sequencing

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Published on: July 5, 2019

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations

Published on: March 29, 2017

Area of Science:

Biotechnology
Molecular Biology
Oncology

Background:

Accurate mutational status is crucial for personalized and cost-effective cancer therapies.
Existing methods like single-gene detection or massive sequencing have limitations.
A need exists for efficient and precise genotyping tools.

Purpose of the Study:

To develop and validate a novel genotyping tool for detecting single-nucleotide variants.
To enable cost-effective and personalized prediction of patient response to therapies.
To provide a rapid and reproducible method for oncological patient discrimination.

Main Methods:

Utilizing a biosensing method with DNA arrays for colorimetric detection.
Employing sequence-tailored probes and PCR with SuperSelective primers for variant discrimination.
Analyzing chip images via fluorescence scanner, documental scanner, or smartphone for spot intensities.

Main Results:

Achieved high discrimination factors in human cell lines with 95% precision and 1% sensitivity for mutant DNA.
Successfully performed selective genotyping of the KRAS gene in tumorous samples (tissue and liquid biopsy).
Corroborated results with Next-Generation Sequencing (NGS) data.

Conclusions:

The developed technology offers a fast, cheap, and reproducible method for discriminating oncological patients.
Low-cost chips and optical reading facilitate the implementation of this genotyping tool.
This approach advances personalized medicine by enabling precise mutational analysis.