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PubMed · 15153623

Expressed sequence tags: medium-throughput protocols.

Claire Whitton·Jennifer Daub·Marian Thompson·Mark Blaxter
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Abstract

Generating expressed sequence tags is a simple, cheap, and efficient way to sample the genome of a target organism. An expressed sequence tag (EST) is a single-pass sequence derived from a single complementary DNA (cDNA) clone, and the sequence serves to identify the gene from which it derives. We present a set of tested laboratory protocols for setting up and performing an EST analysis of any chosen species. These medium-throughput protocols do not require dedicated genomics equipment, such as robots, and focus on the use of microtiter plates and multichannels. Using these protocols, a single competent research worker should be able to generate 2000 ESTs in 1 mo. In a nonnormalized library, these 2000 ESTs should identify between 1000 and 1500 different genes, and thus possibly between 10 and 20% of the genes of any target parasite.

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Base SequenceCloning, MolecularDNA, ComplementaryExpressed Sequence TagsPolymerase Chain ReactionJournal Article

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Claire Whitton, Jennifer Daub, Marian Thompson, Mark Blaxter. 2004. Expressed sequence tags: medium-throughput protocols.. https://doi.org/10.1385/1-59259-793-9%3A075

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