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Related Concept Videos

Next-generation Sequencing03:00

Next-generation Sequencing

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.
Sanger Sequencing01:57

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: May 29, 2026

Integration of Wet and Dry Bench Processes Optimizes Targeted Next-generation Sequencing of Low-quality and Low-quantity Tumor Biopsies
13:24

Integration of Wet and Dry Bench Processes Optimizes Targeted Next-generation Sequencing of Low-quality and Low-quantity Tumor Biopsies

Published on: April 11, 2016

Rapid and efficient human mutation detection using a bench-top next-generation DNA sequencer.

Qian Jiang1, Tychele Turner, Maria X Sosa

  • 1Center for Complex Disease Genomics, McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.

Human Mutation
|September 8, 2011
PubMed
Summary
This summary is machine-generated.

The 454 GS Junior platform efficiently detects mutations in semaphorin genes linked to Hirschsprung disease (HSCR). This rapid next-generation sequencing method identified potential HSCR-causing variants in SEMA3A, SEMA3C, and SEMA3D genes.

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Detection of Rare Mutations in CtDNA Using Next Generation Sequencing
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Detection of Rare Mutations in CtDNA Using Next Generation Sequencing

Published on: August 24, 2017

Related Experiment Videos

Last Updated: May 29, 2026

Integration of Wet and Dry Bench Processes Optimizes Targeted Next-generation Sequencing of Low-quality and Low-quantity Tumor Biopsies
13:24

Integration of Wet and Dry Bench Processes Optimizes Targeted Next-generation Sequencing of Low-quality and Low-quantity Tumor Biopsies

Published on: April 11, 2016

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations
10:41

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

Detection of Rare Mutations in CtDNA Using Next Generation Sequencing
11:11

Detection of Rare Mutations in CtDNA Using Next Generation Sequencing

Published on: August 24, 2017

Area of Science:

  • Genetics and Genomics
  • Molecular Biology
  • Medical Genetics

Background:

  • Next-generation sequencing (NGS) offers high throughput for human mutation detection, enabling simultaneous study of multiple genes and samples with high coverage.
  • Clinical applications often require rapid turnaround times for genetic analyses, particularly for intensive studies of specific genes.

Purpose of the Study:

  • To evaluate the 454 GS Junior platform for mutation detection using amplicon sequencing.
  • To assess its performance for studying type 3 semaphorin genes (SEMA3A, SEMA3C, SEMA3D) associated with Hirschsprung disease (HSCR).

Main Methods:

  • Utilized the bench-top 454 GS Junior platform for amplicon sequencing.
  • Analyzed 39 PCR amplicons (14,014 bp total) from 47 samples, pooled in sets of 12.
  • Sequencing runs averaged ~75,000 reads, ~28 million high-quality bases, with a 371 bp average read length in 10 hours.

Main Results:

  • Achieved an overall sequencing error rate of 0.26 changes per kb at ≥20x coverage depth.
  • Identified 37 sequence variants across the three semaphorin genes.
  • Discovered 10 variants unique to HSCR patients, including five missense mutations potentially linked to HSCR pathogenesis.

Conclusions:

  • The 454 GS Junior platform is a viable solution for rapid, high-coverage mutation detection in targeted gene sets.
  • Identified potential pathogenic mutations in SEMA3A, SEMA3C, and SEMA3D warrant further investigation in larger HSCR patient cohorts.