RT-PCR Protocols

Abstract

Thirty years ago, the investigation of gene sequences and, in particular, disease-causing mutations was a tedious, highly skilled operation; however, this field was revolutionized in the mid-1980s with the introduction and subsequent development of a technique which enabled the reliable amplification of minute quantities of starting DNA into readily detectable levels. Nowadays, this technique, otherwise known as reverse-transcription polymerase chain reaction (RT-PCR), has become routine in most laboratories. Indeed, such is the popularity of RT-PCR that a protocol has been published describing how this could be carried out in a kitchen using common household equipment. The aim of this volume is to translate RT-PCR theory into practice. To achieve this, a comprehensive guide to currently available RT-PCR techniques is given in the form of user-friendly protocols. These protocols contain precise information about all the necessary chemical, consumable, and equipment resources and detailed instructions about how to perform each stage of the different methods. Furthermore, each protocol concludes with a comprehensive notes section, where authors provide helpful hints, trouble-shooting tips, and other must-know information in a format which is accessible to the beginner.