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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.
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RNA-seq03:21

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
DNA Isolation01:24

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...

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Updated: Jul 7, 2026

Pyrosequencing for Microbial Identification and Characterization
12:37

Pyrosequencing for Microbial Identification and Characterization

Published on: August 22, 2013

DNA sequencing by the chemical method.

R L Eckert1

  • 1Case Western Reserve University, Cleveland, Ohio, USA.

Current Protocols in Molecular Biology
|February 12, 2008
PubMed
Summary
This summary is machine-generated.

This protocol details a chemical DNA sequencing method for beginners using radioactive labeling and base-specific modifications. The DNA sequence is determined by analyzing strand breaks on a polyacrylamide gel via autoradiography.

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DamID-seq: Genome-wide Mapping of Protein-DNA Interactions by High Throughput Sequencing of Adenine-methylated DNA Fragments

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Last Updated: Jul 7, 2026

Pyrosequencing for Microbial Identification and Characterization
12:37

Pyrosequencing for Microbial Identification and Characterization

Published on: August 22, 2013

Nanopore DNA Sequencing for Metagenomic Soil Analysis
07:33

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Published on: December 14, 2017

DamID-seq: Genome-wide Mapping of Protein-DNA Interactions by High Throughput Sequencing of Adenine-methylated DNA Fragments
09:14

DamID-seq: Genome-wide Mapping of Protein-DNA Interactions by High Throughput Sequencing of Adenine-methylated DNA Fragments

Published on: January 27, 2016

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Chemical DNA sequencing methods provide essential tools for genetic analysis.
  • Radioactive labeling of DNA fragments is a common prerequisite for sequencing procedures.

Purpose of the Study:

  • To provide a clear, step-by-step protocol for chemical DNA sequencing.
  • To enable researchers with basic molecular biology experience to perform DNA sequencing.

Main Methods:

  • DNA fragments are labeled at one end with radioactive isotopes (32P or 35S).
  • Labeled DNA is divided into four aliquots for base-specific chemical modification.
  • Piperidine is used to induce strand scission at modified bases.
  • Reaction products are separated by denaturing polyacrylamide gel electrophoresis.
  • Autoradiography is performed on the gel to visualize DNA fragments.

Main Results:

  • The method allows for the determination of DNA sequence based on fragment migration patterns.
  • Successful sequencing relies on precise execution of chemical reactions and electrophoresis.

Conclusions:

  • This chemical sequencing protocol is accessible to novice researchers.
  • The method provides a reliable way to determine DNA sequences through specific chemical reactions and gel electrophoresis.