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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.
PCR01:32

PCR

Overview
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...
Real Time RT-PCR02:57

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
PCR - Polymerase Chain Reaction01:32

PCR - Polymerase Chain Reaction

Overview

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Related Experiment Video

Updated: Jun 3, 2026

Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
10:24

Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons

Published on: August 29, 2014

Sequencing of PCR products.

P Moss1, S L Thein

  • 1Institute of Molecular Medicine, John Radcliffe Hospital University of Oxford, UK.

Methods in Molecular Medicine
|March 11, 2011
PubMed
Summary
This summary is machine-generated.

Directly sequencing polymerase chain reaction (PCR) products offers a preferable alternative to traditional subcloning methods. This chapter focuses on optimizing direct DNA sequencing for amplified DNA fragments, enhancing research efficiency.

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Pyrosequencing for Microbial Identification and Characterization
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Pyrosequencing for Microbial Identification and Characterization

Published on: August 22, 2013

Novel Sequence Discovery by Subtractive Genomics
09:40

Novel Sequence Discovery by Subtractive Genomics

Published on: January 25, 2019

Related Experiment Videos

Last Updated: Jun 3, 2026

Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
10:24

Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons

Published on: August 29, 2014

Pyrosequencing for Microbial Identification and Characterization
12:37

Pyrosequencing for Microbial Identification and Characterization

Published on: August 22, 2013

Novel Sequence Discovery by Subtractive Genomics
09:40

Novel Sequence Discovery by Subtractive Genomics

Published on: January 25, 2019

Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • DNA sequencing is crucial after DNA amplification.
  • Conventional methods involve cloning polymerase chain reaction (PCR) products into vectors before sequencing.
  • Direct sequencing of PCR products offers potential advantages.

Purpose of the Study:

  • To address the direct sequencing of PCR products without subcloning.
  • To provide a preferred method for obtaining DNA sequence information from amplified DNA.

Main Methods:

  • Focuses on the direct sequencing approach for PCR products.
  • Details methodologies for sequencing amplified DNA fragments directly.

Main Results:

  • Direct sequencing eliminates the need for intermediate cloning steps.
  • This method can be more efficient than traditional subcloning and sequencing.

Conclusions:

  • Direct sequencing of PCR products is a valuable and often preferable technique.
  • Optimizing this approach streamlines the process of obtaining DNA sequence data.