Expressed Sequence Tags (ESTs)

Leading experts in the field introduce the reader to many of the fundamental concepts underlying the generation and analysis of ESTs through accessible and affordable sequencing technologies. The book includes reproducible protocols and troubleshooting advice.

Over the past two decades, expressed sequence tags (ESTs - single pass reads from randomly selected cDNAs), have proven to be a remarkably cost-effective route for the purposes of gene discovery. Gaining in popularity, millions of ESTs have now been generated for over a thousand different species. In Expressed Sequence Tags (ESTs): Generation and Analysis , leading experts in the field introduce the reader to many of the fundamental concepts underlying the generation and analysis of ESTs through readily accessible and affordable sequencing technologies. The volume focuses on various methods used to generate, process and analyze EST datasets, while also exploring the use of EST technology for other purposes such as expression profiling, analysis of alternative transcripts, and phylogenomics. Written in the highly successful Methods in Molecular Biology™ series format, chapters include brief introductions to their respective topics, lists of the necessary materials and programs, step-by-step, readily reproducible protocols, and Notes sections, which highlight tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and easy to use, Expressed Sequence Tags (ESTs): Generation and Analysis serves as an ideal reference for scientists continuing the vital investigation of genes and the genome.

Essential reading for any scientist beginning their own EST project Takes the reader through the entire process of how to successfully complete an EST project Covers a wide range of cDNA library construction techniques from the generation of full length libraries to the use of splice leaders and normalization and subtraction procedures Details state-of-the-art bioinformatics pipelines and tools that can be readily implemented for high throughput processing and analysis of ESTs Shows the reader how EST technology can be exploited for expression studies, particularly from the perspective of human health Provides strategies, tips and potential pitfalls in the generation and analysis of ESTs from leading exponents of the field Includes supplementary material: sn.pub/extras

Inhalt
Expressed Sequence Tags: An Overview.- Strategies for Undertaking Expressed Sequence Tag (EST) Projects.- Construction of cDNA Libraries: Focus on Protists and Fungi.- Generation of Full-Length cDNA Libraries: Focus on Plants.- Preparation of Full-Length cDNA Libraries: Focus on Metazoans.- Expressed Sequence Tags: Normalization and Subtraction of cDNA Libraries Expressed sequence tags normalization and subtraction of cDNA libraries .- Generating EST Libraries: Trans-Spliced cDNAs.- Large-Scale Sequencing and Analytical Processing of ESTs.- EST Processing: From Trace to Sequence.- Obtaining Accurate Translations from Expressed Sequence Tags.- EST Databases and Web Tools for EST Projects.- Phylogenomic Analysis of EST Datasets.- Statistical Analysis of Expressed Sequence Tags.- Application of ESTs in Microarray Analysis.- Exploiting ESTs in Human Health.