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Related Concept Videos

Sanger Sequencing01:57

Sanger Sequencing

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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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Next-generation Sequencing03:00

Next-generation Sequencing

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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.
Next-Generation Sequencing Methods
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RNA-seq03:21

RNA-seq

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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. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
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Genomics02:02

Genomics

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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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Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

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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.
Challenges of the Maxam-Gilbert Method
The...
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Genome Annotation and Assembly03:36

Genome Annotation and Assembly

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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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Ultra-long Read Sequencing for Whole Genomic DNA Analysis
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Ultra-long Read Sequencing for Whole Genomic DNA Analysis

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Genome Sequencing.

Mansi Verma1, Samarth Kulshrestha2, Ayush Puri2

  • 1Sri Venkateswara College, University of Delhi (South Campus), Benito Juarez Road, Dhaula Kuan, New Delhi, 110 021, India. mansiverma20@gmail.com.

Methods in Molecular Biology (Clifton, N.J.)
|November 30, 2016
PubMed
Summary
This summary is machine-generated.

Genome sequencing advances genotype-phenotype correlation. This review covers Sanger sequencing, Next-Generation Sequencing (NGS), and future real-time techniques for genome analysis.

Keywords:
Cyclic-array sequencingNanoporeNext-generation sequencingSanger sequencing

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Area of Science:

  • Genomics
  • Molecular Biology
  • Bioinformatics

Background:

  • Genome sequencing is crucial for understanding genotype-phenotype relationships across diverse life science fields.
  • Sequencing technologies have evolved through three generations, starting with Sanger and Maxam-Gilbert methods.
  • Next-Generation Sequencing (NGS) offers speed and cost benefits but often lacks the accuracy and read length of Sanger sequencing.

Purpose of the Study:

  • To review available genome sequencing technologies.
  • To provide a generic workflow for genome sequencing.
  • To highlight the ongoing need for improved real-time sequencing techniques.

Main Methods:

  • Review of first-generation sequencing (Sanger, Maxam-Gilbert).
  • Discussion of Next-Generation Sequencing (NGS) technologies (second and third generation).
  • Exploration of emerging real-time sequencing techniques.

Main Results:

  • Sanger sequencing remains a benchmark for accuracy and read length.
  • NGS excels in speed, cost-efficiency, and parallelism, primarily for resequencing.
  • Limitations in NGS accuracy and read length necessitate further technological development.

Conclusions:

  • Continued innovation in sequencing technology is vital for advancing genomic research.
  • Real-time sequencing methods are essential for overcoming current limitations.
  • A comprehensive understanding of sequencing workflows aids genome analysis.