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The in situ hybridization and PCR technologies are now well-established molecular techniques for studying chromosomal aneuploidy and rearran- ments, gene localization and expression, and genomic organization. Over the last decade, we have seen increasing applications in these fields. By combining the high sensitivity of the PCR reaction and the cytological localization of target sequences, both PRINS and in situ PCR techniques have provided highly powerful complements to FISH for in situ cellular and molecular investigations. Both these approaches have several advantages in terms of sensitivity and specificity, owing to the use of primers and to the fast kinetics of annealing and elongation reactions in situ. In the first edition of PRINS and In Situ PCR Protocols edited by John R. Gosden, experts in the field presented in detail a variety of applications of PRINS and in situ PCR techniques, in a wide range of clinical conditions. Since the publication of this successful reference book, there have been s- nificant improvements in in situ detection techniques. This completely revised and updated second edition presents a compreh- sive selection of new procedures developed in the field of PRINS and in situ PCR technologies. The book has two sections. Part I, Basic Methodology, contains chapters that provide useful protocols for many variations of PRINS and in situ PCR, including a new fast multicolor PRINS method, and protocols for PRINS detection of unique sequences in situ.

PRINS and In Situ PCR Protocols

Part I. Basic Methodology

New Rapid Multicolor PRINS Protocol
Ju Yan, Macoura Gadji, Kada Krabchi, and Régen Drouin

PRINS Combined With Peptide Nucleic Acid Labeling
Franck Pellestor and Petra Paulasova

PRINS Combined With Indirect Immunofluorescence
Philippe Coullin

PRINS for the Detection of Unique Sequences
Stephen S. Wachtel and Avirachan T. Tharapel

Single-Copy and Point Mutation Analysis (Rolling-Circle PRINS)
Jorn Koch

Analysis of Sperm Aneuploidy by PRINS
Franck Pellestor, Brigitte Andréo, Jacques Puechberty, Genevieve Lefort, and Pierre Sarda

In Situ PCR Protocols
Omar Bagasra and Twaina Harris

Part II. Research and Clinical Applications

Detection and Sizing of Telomeric and Other Simple Repeats by Dideoxy-PRINS
Jorn Koch

PRINS Evaluation of Chromosome Instability in Mammalian Cells by Detection of Repetitive DNA Sequences in Micronuclei
Antonella Russo

PRINS for the Detection of Gene Deletions in Cancer
Avirachan T. Tharapel and Stephen S. Wachtel

Combined Fluorescence In Situ Hybridization and PRINS for the Analysis of the Dystrophin Gene
Liborio Stuppia, Dario La Sala, and Caterina Cinti

Brain Tissue Preparations for Chromosomal PRINS Labeling
Ivan Y. Iourov, Svetlana G. Vorsanova, Franck Pellestor, and Yuri B. Yurov

PRINS on Plant Chromosomes
Jirí Macas, Alice Navrátilová, Marie Kubaláková, and Jaroslav Dolezel

Dual-Color PRINS for In Situ Detection of Fetal Cells in Maternal Blood
Kada Krabchi, Macoura Gadji, Ju Yan, and Régen Drouin

PRINS as an Efficient Tool for Aneuploidy Assessment in Human Oocytes and Preimplantation Embryos
Franck Pellestor, Brigitte Andréo, Tal Anahory, and Samir Hamamah

Microdissection With PCR In Situ
Allen T. Christian and ChristineHara

Application of Reverse Transcription In Situ PCR in Cancer Analysis
Tamotsu Takeuchi, Yoshihiro Adachi, Hiroshi Sonobe, and Yuji Ohtsuki

In Situ Reverse Transcription PCR on Plant Tissues
Zbigniew Przybecki, Ewa Siedlecka, Marcin Filipecki, and Ewa Urbanczyk-Wochniak

In Situ PCR for the Detection of Human Cytomegalovirus in Suspension Cells During the Latent Phase of Infection
Barry Slobedman

Development and Adaptation of the PRINS Technology: An Overview
Franck Pellestor

Latest Developments in In Situ PCR
Omar Bagasra and Twaina Harris

Index