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Related Concept Videos

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...
Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...
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.
Mismatch Repair01:36

Mismatch Repair

Overview
Mismatch Repair01:20

Mismatch Repair

Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...

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Related Experiment Video

Updated: Jul 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 by cycle sequencing.

L Thierfelder1

  • 1Max-Delbrück-Centrum für Molekulare Medizin, Berlin, Germany.

Current Protocols in Human Genetics
|April 23, 2008
PubMed
Summary
This summary is machine-generated.

Cycle sequencing simplifies identifying gene mutations by eliminating the need for subcloning. This DNA sequencing method allows screening of PCR products for mutations in candidate genes.

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The Lambda Select cII Mutation Detection System
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The Lambda Select cII Mutation Detection System

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Related Experiment Videos

Last Updated: Jul 5, 2026

Rare Event Detection Using Error-corrected DNA and RNA Sequencing
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Published on: August 3, 2018

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

The Lambda Select cII Mutation Detection System
07:08

The Lambda Select cII Mutation Detection System

Published on: April 26, 2018

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Screening candidate genes for mutations is crucial for understanding genetic diseases.
  • Conventional DNA sequencing methods can be laborious and time-consuming, often requiring subcloning steps.

Purpose of the Study:

  • To describe and evaluate cycle sequencing as a simplified method for mutation detection in candidate genes.
  • To highlight the advantages of cycle sequencing over traditional sequencing techniques for genetic analysis.

Main Methods:

  • Polymerase Chain Reaction (PCR) amplification of candidate gene segments from genomic DNA.
  • Multiple rounds of amplification using a thermal cycler, heat-stable DNA polymerase, dideoxynucleotides, and a radiolabeled primer.
  • Fractionation of 32P-labeled sequencing products on denaturing polyacrylamide gels, followed by autoradiography.

Main Results:

  • Cycle sequencing enables the screening of approximately 200 base pairs of DNA from 10 to 30 individuals per gel.
  • The method effectively identifies mutations without requiring the subcloning of genomic fragments or PCR products.
  • This technique offers a simpler and more efficient approach compared to conventional sequencing.

Conclusions:

  • Cycle sequencing is a significantly simpler and more efficient method for identifying mutations in candidate genes.
  • The elimination of subcloning steps makes this DNA sequencing technique more accessible for genetic research.
  • Cycle sequencing advances the field of molecular diagnostics and genetic mutation screening.