The Basics of RT-PCR: Some Practical Considerations

The basic reverse-transcriptase-polymerase chain reaction (RT-PCR) technique is used routinely and widely in most biomedical research laboratories. The fundamental considerations for such basics as primer design, good laboratory set up and practice, and techniques for performing RT-PCR have been fully examined elsewhere (1 ,2 ). Thus, I will include only observations from my own experience in this chapter. Although the purpose of this chapter is to help the reader in setting up their own PCR assays, and to provide tips on effective PCR laboratory set up, it should be noted that each individual chapter in this volume contains complete experimental detail on the protocols described.