PCR 3

PCR in situ hybridization allows the detection of specific nucleic acid sequences and their distribution in the cell. It combines two powerful techniquesin situ hybridization (ISH), which allows cellular localization of DNA and RNA sequences in cells and tissues, and the polymerase chain reaction (PCR), which allows reproducible amplification of rare nucleic acid sequences. The combined technique and its variants greatly enhance the sensitivity of in situ hybridization and add morphological localization to the sensitivity of PCR. Such techniques have enormous potential for research and diagnosis but problems with reproducibility and reliability are often encountered. This book overcomes these problems by describing the key procedures in step-by-step detail and by providing the essential advice needed for success. Topics include: DNA in situ PCR and DNA PCR in situ hybridization (PCR ISH) for the detection of DNA targets in cells; reverse trancriptase in situ PCR (RT-PCR) and RT-PCR ISH for the detection of RNA targets; and PRINS (primed in situ synthesis) for chromosomal analysis in interphase nuclei and metaphase chromosome spreads. There are further chapters on fixation of tissues for PCR, selective ultraviolet radiation fractionation (SURF), application of in situ PCR to human tissues, applications and modifications of PCR-ISH, and automation of in situ amplification. PCR In Situ Hybridization is a unique and timely collection of well-tested protocols for the amplification of DNA and RNA in cells and tissues, drawing on the accumulated knowledge and experience of leading exponents of these techniques. For each topic covered, the authors provide detailed guidance on the key steps in the protocols, numerous hints and tips for success, and advice on trouble-shooting. PCR In Situ Hybridization will be invaluable to molecular biologists, pathologists, geneticists, and all those seeking to perform in situ analyses of nucleic acid molecules.

Readership: Graduates and post-doctoral researchers, academic staff, and clinicians across a wide range of molecular medical disciplines.

Edited by C. Simon Herrington, Senior Lecturer, University of Liverpool, and John J. O'Leary, Doctor, John Radcliffe Hospital, Oxford

""...the book gives an excellent overview of the practical application of PCRIS." Molecular Pathology, 1999"

""This is one of an impressive series of laboratory handbooks entitled The Practical Approach Series...The presentation of this material is of uniformly high quality...this book would be a valuable resource in a lab" Canadian Journal of Medical Laboratory Scince"

1: Fixation of tissues for the polymerase chain reaction
2: In situ genetic analysis with selective ultraviolet radiation fractionation (SURF)
3: In situ hybridization
4: PCR in situ hybridization (PCR-ISH)
5: Reverse transcriptase in situ PCR for RNA detection
6: Primed in situ DNA synthesis (PRINS)
7: Application of in situ-PCR techniques to human tissues
8: Applications and modifications of PCR in situ hybridization
9: Automation of in situ amplification