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(TM) 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