Specific Synthesis of Dna Invitro Via A Polymerase-Catalyzed Chain-Reaction

This chapter discusses specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. A source of DNA including the desired sequence is denatured in the presence of a large molar excess of two oligonucleotides and the four deoxyribonucleoside triphosphates. Oligonucleotides were synthesized using an automated DNA synthesis machine using phosphoramidite chemistry. Mispriming can be usefully employed to make intentional in vitro mutations or to add sequence information to one or both ends of a given sequence. Amplifications of other human loci have resulted in varying degrees of specificity and efficiency. The polymerase chain reaction has thus found immediate use in developmental DNA diagnostic procedures and in molecular cloning from genomic DNA.