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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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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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Related Experiment Video

Updated: Dec 11, 2025

Amplification of Near Full-length HIV-1 Proviruses for Next-Generation Sequencing
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Highly Multiplexed, Semiautomated Nextera Next-Generation Sequencing (NGS) Library Preparation.

William Christie1, Ron Yadin1, Kristy Ip1

  • 1Amyris Inc., Emeryville, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|August 19, 2020
PubMed
Summary

This study presents a high-throughput DNA sequencing protocol using miniaturized Nextera Tagmentation reactions and custom PCR index primers. This method significantly reduces costs for sequencing thousands of DNA constructs in a single run.

Keywords:
High-throughputMiniaturizationMultiplexed next-generation sequencingSynthetic biology

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • High-throughput DNA sequencing is crucial for large-scale DNA assembly.
  • Current methods can be costly and time-consuming for massive projects.

Purpose of the Study:

  • To develop a cost-effective, high-throughput protocol for DNA sequencing.
  • To enable simultaneous sequencing of thousands of DNA constructs.

Main Methods:

  • Utilized traditional and acoustic liquid-handling robotics for miniaturization.
  • Employed Illumina's Nextera Tagmentation reactions with custom PCR index primers.
  • Generated highly multiplexed next-generation sequencing (NGS) libraries for pooled sequencing.

Main Results:

  • Successfully miniaturized Nextera Tagmentation reactions.
  • Enabled the production of highly multiplexed NGS libraries.
  • Achieved simultaneous sequencing of thousands of DNA constructs in a single run.

Conclusions:

  • The described protocol offers a dramatically reduced cost for high-throughput DNA sequencing.
  • This method is suitable for large-scale DNA assembly operations requiring pooled sequencing.