The Basics of RT-PCR

Some Practical Considerations
  • Joe O’Connell
Part of the Methods in Molecular Biology book series (MIMB, volume 193)


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.


Optimal Annealing Temperature Guanidine Thiocyanate Household Bleach Nonspecific Priming Cesium Chloride Density Gradient 
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Copyright information

© Humana Press Inc. 2002

Authors and Affiliations

  • Joe O’Connell
    • 1
  1. 1.Department of MedicineNational University of IrelandCorkIreland

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