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Related Concept Videos

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
Although all next-generation methods use different technologies, they all share a set of standard features....
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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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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. 
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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.
Challenges of the Maxam-Gilbert Method
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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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Related Experiment Video

Updated: May 3, 2026

Targeted Next-generation Sequencing and Bioinformatics Pipeline to Evaluate Genetic Determinants of Constitutional Disease
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Library construction for next-generation sequencing: overviews and challenges.

Steven R Head1, H Kiyomi Komori2, Sarah A LaMere2

  • 1NGS and Microarray Core Facility, The Scripps Research Institute, La Jolla, CA.

Biotechniques
|February 8, 2014
PubMed
Summary
This summary is machine-generated.

High-throughput sequencing, or next-generation sequencing (NGS), offers powerful genomic insights. This review focuses on optimizing NGS library preparation from DNA and RNA for diverse applications, ensuring high-quality results.

Keywords:
ChIP-seqDNADNA-seqRIP-seqRNARNA-seqdeep sequencinglibrary preparationnext-generation sequencing

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Area of Science:

  • Genomics
  • Molecular Biology
  • Bioinformatics

Background:

  • High-throughput sequencing (NGS) has transformed genomic research.
  • NGS technology continues to advance, reducing costs and expanding applications.
  • The quality of sequencing libraries is crucial for reliable genomic data.

Purpose of the Study:

  • To examine the critical role of sequencing library quality in NGS.
  • To discuss challenges in preparing NGS libraries from DNA and RNA.
  • To cover preparation methods for various NGS applications and single-cell sequencing.

Main Methods:

  • Review of factors influencing DNA and RNA source material for library preparation.
  • Analysis of considerations for diverse NGS applications (genome, targeted, RNA-seq, ChIP-seq, RIP-seq, methylation).
  • Discussion of current methodologies for single-cell NGS library preparation.

Main Results:

  • Library quality is paramount for successful NGS outcomes.
  • Source material quantity and characteristics significantly impact library preparation.
  • Application-specific requirements necessitate tailored library preparation strategies.

Conclusions:

  • Optimizing NGS library preparation is key to maximizing data quality and research potential.
  • Addressing challenges in DNA/RNA input and application-specific needs is essential.
  • Advancements in single-cell library preparation enable deeper genomic insights.