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Related Concept Videos

Sanger Sequencing01:57

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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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RNA-seq03:21

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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. 
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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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Genomics02:02

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Genomics is the science of genomes: it is the study of all the genetic material of an organism. In humans, the genome consists of information carried in 23 pairs of chromosomes in the nucleus, as well as mitochondrial DNA. In genomics, both coding and non-coding DNA is sequenced and analyzed. Genomics allows a better understanding of all living things, their evolution, and their diversity. It has a myriad of uses: for example, to build phylogenetic trees, to improve productivity and...
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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.
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The genome refers to all of the genetic material in an organism. It can range from a few million base pairs in microbial cells to several billion base pairs in many eukaryotic organisms. Genome assembly refers to the process of taking the DNA sequencing data and putting it all back together in a correct order to create a close representation of the original genome. This is followed by the identification of functional elements on the newly assembled genome, a process called genome annotation.
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Rare Event Detection Using Error-corrected DNA and RNA Sequencing
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Exoquence DNA sequencing

C Li1, P W Tucker

  • 1Department of Microbiology, University of Texas Southwestern Medical Center, Dallas 75235.

Nucleic Acids Research
|March 11, 1993
PubMed
Summary
This summary is machine-generated.

This new DNA sequencing strategy uses specific enzyme digestion for efficient template preparation, enabling rapid and high-quality sequencing without prior DNA knowledge or subcloning.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Traditional DNA sequencing methods often require subcloning and prior knowledge of the DNA sequence.
  • The preparation of sequencing templates can be time-consuming and complex.

Purpose of the Study:

  • To develop a streamlined and efficient strategy for DNA sequencing.
  • To eliminate the need for subcloning and oligonucleotide primers in DNA sequencing.
  • To enable high-quality sequencing results from uncharacterized DNA samples.

Main Methods:

  • Utilized exonuclease III digestion followed by double-strand specific endonuclease digestion.
  • Employed direct dideoxynucleotide sequencing reactions.
  • Generated single-stranded DNA templates and primers from double-stranded DNA.

Main Results:

  • Successfully developed a strategy that bypasses the need for subcloning and primers.
  • Demonstrated the ability to prepare sequencing templates from uncharacterized DNA in one day.
  • Achieved high-quality sequencing results suitable for automated and routine laboratory use.
  • Enabled simultaneous sequence information retrieval from multiple DNA regions.

Conclusions:

  • The developed DNA sequencing strategy is efficient, rapid, and versatile.
  • This method simplifies the sequencing process, making it accessible for uncharacterized DNA.
  • The approach is compatible with existing dideoxy-sequencing kits and automation.