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Related Concept Videos

Next-generation Sequencing03:00

Next-generation Sequencing

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.
Sanger Sequencing01:57

Sanger Sequencing

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...
RNA-seq03:21

RNA-seq

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 microarray-based...
Real Time RT-PCR02:57

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

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Related Experiment Video

Updated: May 8, 2026

Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
10:24

Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons

Published on: August 29, 2014

Isothermal amplification method for next-generation sequencing.

Zhaochun Ma1, Raymond W Lee, Bin Li

  • 1Genetic Systems, Life Technologies Inc., South San Francisco, CA 94080, USA. zhaochun.ma@lifetech.com

Proceedings of the National Academy of Sciences of the United States of America
|August 14, 2013
PubMed
Summary
This summary is machine-generated.

We developed a novel isothermal amplification method to create immobilized DNA templates for next-generation sequencing. This technique generates over a billion colonies on a sequencing flowchip, advancing high-throughput sequencing applications.

Keywords:
DNA breathingforming coloniesgenomein situ PCRpolony

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Last Updated: May 8, 2026

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Pre-Implantation Genetic Testing for Aneuploidy on a Semiconductor Based Next-Generation Sequencing Platform

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genomics

Background:

  • Next-generation sequencing (NGS) requires high-quality DNA templates for efficient data generation.
  • Current methods for template preparation can be complex and limit throughput.

Purpose of the Study:

  • To develop an efficient method for generating immobilized monoclonal DNA templates for NGS.
  • To enhance the scalability and efficiency of template preparation for high-throughput sequencing.

Main Methods:

  • An isothermal amplification approach utilizing a template walking mechanism.
  • Employing low-melting temperature (Tm) solid-surface homopolymer primers and a low-Tm solution phase primer.
  • Demonstration of an alternative paired-end sequencing method using interstrand DNA photo cross-linking.

Main Results:

  • Generation of over one billion submicrometer-sized colonies in a single NGS flowchip lane.
  • Successful immobilization of monoclonal templates on a solid surface.
  • Demonstration of a novel photo cross-linking technique for template linkage.

Conclusions:

  • The described isothermal amplification method provides a scalable and efficient way to generate immobilized templates for NGS.
  • This approach has the potential to significantly improve the throughput and cost-effectiveness of next-generation sequencing applications.
  • The alternative photo cross-linking method offers a new strategy for paired-end sequencing template preparation.