High quality cDNA libraries for discovery and validation of novel drug targets

Abstract

High quality, full-length cDNA libraries are essential for a successful functional genomics approach to discover and validate novel drug targets. This article reviews various techniques and strategies for construction of high quality, full-length cDNA libraries, which should have better preservation of mRNA-derived sequences and be devoid of contaminating sequences and cloning artefacts. Key elements in every step of cDNA library construction, including RNA isolation, first- and second-strand cDNA synthesis and assembly into cloning vectors are pinpointed to avoid pitfalls, increase the full-length cDNA ratio and eliminate undesirable clones in the final library.

• 5-directed
• 5-enriched
• cap structure
• cDNA libraries
• first-strand cDNA
• full-length cDNA
• functional genomics
• mRNA
• reverse transcriptase
• second-strand cDNA
• vector-primer