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Related Concept Videos

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...
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...

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Related Experiment Video

Updated: Jun 24, 2026

High-throughput Identification of Gene Regulatory Sequences Using Next-generation Sequencing of Circular Chromosome Conformation Capture (4C-seq)
09:06

High-throughput Identification of Gene Regulatory Sequences Using Next-generation Sequencing of Circular Chromosome Conformation Capture (4C-seq)

Published on: October 5, 2018

Large-scale sequencing and analytical processing of ESTs.

Makedonka Mitreva1, Elaine R Mardis

  • 1Genome Sequencing Center, Department of Genetics, Washington University School of Medicine, St. Louis, MO, USA.

Methods in Molecular Biology (Clifton, N.J.)
|March 12, 2009
PubMed
Summary
This summary is machine-generated.

Expressed sequence tags (ESTs) offer rapid gene discovery in eukaryotes. Large-scale sequencing on ABI 3730xl and 454 GS20 platforms, coupled with bioinformatics, enables comprehensive transcriptome exploration.

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3' End Sequencing Library Preparation with A-seq2
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Last Updated: Jun 24, 2026

High-throughput Identification of Gene Regulatory Sequences Using Next-generation Sequencing of Circular Chromosome Conformation Capture (4C-seq)
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3' End Sequencing Library Preparation with A-seq2
12:01

3' End Sequencing Library Preparation with A-seq2

Published on: October 10, 2017

Area of Science:

  • Genomics
  • Molecular Biology
  • Bioinformatics

Background:

  • Expressed sequence tags (ESTs) are crucial for rapid gene discovery in eukaryotic genomes.
  • ESTs have diverse applications including probe design, alternative splicing analysis, and gene boundary verification.
  • Increasing EST data in public databases like GenBank reflects advancements in molecular techniques and sequencing technologies.

Purpose of the Study:

  • To describe large-scale expressed sequence tag (EST) sequencing on distinct platforms.
  • To detail the processes of sequence extraction, processing, and submission to public databases.
  • To present methods for generating full-length cDNA sequences using 454 GS20 pyrosequencing.

Main Methods:

  • Large-scale EST sequencing utilizing ABI 3730xl and 454 Life Sciences GS20 platforms.
  • Conventional Sanger sequencing from existing cDNA libraries.
  • 454 GS20 pyrosequencing, including a method for full-length cDNA generation.
  • Sequence data processing and submission to public repositories.

Main Results:

  • Demonstration of large-scale EST sequencing capabilities on two different platforms.
  • Successful extraction, processing, and submission of sequence data.
  • Integration of conventional and pyrosequencing methods for comprehensive transcriptome analysis.

Conclusions:

  • Both ABI 3730xl and 454 GS20 platforms are effective for large-scale EST sequencing.
  • Combining these platforms with bioinformatics tools provides a powerful approach for exploring organism transcriptomes.
  • Advancements in sequencing technology continue to enhance the utility and scope of ESTs in genomic research.