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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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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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Maxam-Gilbert Sequencing01:05

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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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Evolutionary Relationships through Genome Comparisons02:54

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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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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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Sequencing of mRNA from Whole Blood using Nanopore Sequencing
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Evolution of DNA sequencing.

Hamid Nawaz Tipu1, Ambreen Shabbir2

  • 1Department of Pathology, Combined Military Hospital, Khuzdar Cantt.

Journal of the College of Physicians and Surgeons--Pakistan : JCPSP
|March 17, 2015
PubMed
Summary
This summary is machine-generated.

Sanger DNA sequencing, though foundational, has limitations. Next Generation Sequencing (NGS) technologies offer improved sensitivity and broader applications in clinical diagnostics and research.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • The advent of Sanger DNA sequencing in the 1970s revolutionized genetic analysis.
  • Original Sanger sequencing relied on dideoxythymidine triphosphate (ddTTP) chain termination and radiographic detection via Polyacrylamide Gel Electrophoresis (PAGE).
  • Subsequent improvements included fluorescence detection, capillary electrophoresis, and capillary array electrophoresis.

Purpose of the Study:

  • To review the evolution of DNA sequencing technologies.
  • To highlight the limitations of Sanger sequencing.
  • To introduce and discuss the impact of Next Generation Sequencing (NGS) technologies.

Main Methods:

  • Review of historical DNA sequencing methodologies.
  • Discussion of advancements in Sanger sequencing techniques.
  • Overview of Next Generation Sequencing (NGS) platforms and their capabilities.

Main Results:

  • Sanger sequencing faced limitations including decreased sensitivity for low-level mutations, challenges with highly polymorphic regions (e.g., Major Histocompatibility Complex), and high DNA concentration requirements.
  • Next Generation Sequencing (NGS) technologies have been developed to overcome these limitations.
  • NGS enables applications such as cancer variant studies, minimal residual disease detection, exome sequencing, SNP analysis, epigenetics, and microbial genome sequencing.

Conclusions:

  • NGS technologies represent a significant advancement over traditional Sanger sequencing.
  • The integration of NGS into clinical research and diagnostics is poised to transform medical approaches.
  • NGS offers enhanced capabilities for a wide range of genetic and genomic analyses.