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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...
Mismatch Repair01:36

Mismatch Repair

Overview

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Related Experiment Video

Updated: Jun 5, 2026

Rare Event Detection Using Error-corrected DNA and RNA Sequencing
10:36

Rare Event Detection Using Error-corrected DNA and RNA Sequencing

Published on: August 3, 2018

Mutation detection with next-generation resequencing through a mediator genome.

Omri Wurtzel1, Mally Dori-Bachash, Shmuel Pietrokovski

  • 1Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.

Plos One
|January 7, 2011
PubMed
Summary

Next-generation sequencing (NGS) enables bacterial mutation analysis. This new framework uses related genomes as mediators, overcoming the need for a wild-type reference genome to identify causative mutations.

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Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter
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Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter

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Related Experiment Videos

Last Updated: Jun 5, 2026

Rare Event Detection Using Error-corrected DNA and RNA Sequencing
10:36

Rare Event Detection Using Error-corrected DNA and RNA Sequencing

Published on: August 3, 2018

Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter
06:59

Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter

Published on: March 31, 2022

Area of Science:

  • Microbiology
  • Genomics
  • Bioinformatics

Background:

  • Next-generation sequencing (NGS) is revolutionizing bacterial mutation analysis.
  • Identifying the genetic basis of phenotypic changes requires comparing mutant and wild-type (WT) genomes.
  • A key limitation is the requirement for an existing WT reference genome for short-read sequencing data, hindering de novo assembly.

Purpose of the Study:

  • To develop a general framework for bacterial mutation analysis using NGS without a specific WT reference genome.
  • To enable accurate identification of causative mutations by leveraging related species' genomes as mediators.

Main Methods:

  • Sequencing of mutant and WT bacterial genomes using NGS.
  • Mapping short sequencing reads to a mediator genome from a related organism.
  • Filtering out variations present in both mutant and WT compared to the mediator to isolate true differences.

Main Results:

  • The developed framework successfully identified a single causative mutation in a prey-independent mutant of *Bdellovibrio bacteriovorus* 109J, despite significant genomic differences with the mediator species (*B. bacteriovorus* HD100).
  • Experimental validation confirmed the identified gene in 53 additional mutants.
  • The approach demonstrated high accuracy in pinpointing causative mutations.

Conclusions:

  • This novel framework significantly expands the utility of NGS for bacterial mutant analysis.
  • It removes the dependency on pre-existing reference genomes, making mutation discovery more accessible.
  • The method provides a robust solution for identifying genetic alterations in bacteria without a direct WT genome sequence.