An efficient vector-primer cDNA cloning system.

Publisher Summary This chapter discusses the efficient vector-primer complementary deoxyribo nucleic acid (cDNA) cloning system. The dimer-primer method allows vector-primed cDNA synthesis from both ends of a T-tailed plasmid vector. Because the starting plasmid approximates a head-to-tail dimer, both cDNA are attached to a functional plasmid bearing a selectable marker gene. This chapter also presents the cDNA synthesis results in DNA copies of two messenger ribo nucleic acid (mRNA) molecules, each covalently attached to one end of the dimer-primer. Vector-primed cDNA cloning has several advantages over other cDNA cloning methods. These include (1) much higher efficiency than the older methods of cloning double-stranded cDNA into a plasmid; (2) high recovery of full-length cDNA; (3) it is simple, fast, and reliable, once a supply of vector and linker have been prepared and tested; and (4) analysis and recovery of the inserts is much easier than in phage cDNA cloning methods, which it now rivals in efficiency.