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Updated: Jun 24, 2026

3' End Sequencing Library Preparation with A-seq2

3' End Sequencing Library Preparation with A-seq2

Published on: October 10, 2017

Strategies for undertaking expressed sequence tag (EST) projects.

Sandra W Clifton¹, Makedonka Mitreva

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

Summary

This summary is machine-generated.

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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 microarray-based...

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Complementary DNA (cDNA) sequencing provides valuable expressed sequence tag (EST) data for genome annotation and gene discovery. Careful planning of project goals, library construction, and data analysis is crucial for successful cDNA sequencing projects.

Area of Science:

Genomics
Molecular Biology
Bioinformatics

Background:

Complementary DNA (cDNA) sequencing is a powerful technique for sampling an organism's transcriptome.
Generated expressed sequence tag (EST) sequences are vital for enhancing genome utility and creating gene catalogs, especially for unsequenced genomes.

Purpose of the Study:

To outline the critical criteria for planning and executing successful cDNA sequencing projects.
To guide researchers in optimizing library construction and data analysis strategies.

Main Methods:

Consideration of project-specific factors: organism choice, tissue selection and pooling, tissue quality, and library type.
Strategic library construction to meet project requirements, with options for outsourcing if needed.
Awareness of emerging sequencing technologies and necessary bioinformatics support.

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An Experimental and Bioinformatics Protocol for RNA-seq Analyses of Photoperiodic Diapause in the Asian Tiger Mosquito, Aedes albopictus

An Experimental and Bioinformatics Protocol for RNA-seq Analyses of Photoperiodic Diapause in the Asian Tiger Mosquito, Aedes albopictus

Published on: November 30, 2014

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Last Updated: Jun 24, 2026

3' End Sequencing Library Preparation with A-seq2

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An Experimental and Bioinformatics Protocol for RNA-seq Analyses of Photoperiodic Diapause in the Asian Tiger Mosquito, Aedes albopictus

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Main Results:

EST data can significantly aid in genome annotation and gene discovery.
Careful consideration of experimental design, including tissue choice and library type, is paramount.
Newer sequencing platforms offer potential for faster and more economical cDNA data generation.

Conclusions:

Successful cDNA sequencing requires meticulous planning, appropriate library construction, and robust data analysis.
Researchers should consider outsourcing specialized tasks and stay informed about technological advancements.
Securing adequate funding through grants, private sources, or collaborations is essential for project completion.