ABSTRACT
This chapter discusses an efficient approach by which representative full-length coding sequences and clones for longer mammalian gene transcripts may be obtained. The genomic revolution manifested by completion of human and other mammalian genome sequences and the ready public access to complementary deoxyribonucleic acid (cDNA) libraries, sequences, and clones will impact greatly on conceptual approaches and experimental strategies for our understanding of nervous system function and complex behaviors The full-length cDNA serves as effective material for functional analysis of a gene, e.g., recombinant protein synthesis, protein characterization of newly isolated genes, generation of knockout mice, and expression profiling by microarray analysis. In order to maximize the yield and fidelity of the oligo capping, complete recovery of the aqueous layer after phenol: chloroform extraction should not be attempted; proteins, lipids and carbohydrates often located in the intermediate layer as floating particles will inhibit the subsequent enzymatic reactions.
