Humana PRINS and In Situ PCR Protocols: 334 (Methods in Molecular Biology, 334)

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. Review From Reviews of the First Edition... "...essential for any laboratory contemplating exploiting this powerful technique." -Microbiology Today "...[a] splendid little book which covers the field quite well,,," -FEBS Letters "...accessible and very practical...recommend[ed]...to any laboratory with an operational PCR machine...a valuable addition to the methods literature." -Quarterly Review of Biology "...well written...contains sufficient detail for a novice to successfully complete the experiment...recommend[ed]...for libraries as well as individual researchers interested in applying PRINS technology." -Doody's Health Sciences Book Review Journal From the Back Cover Carrying on the high standards of the much-praised first edition, leading researchers and clinical investigators have completely revised and updated PRINS and In Situ PCR Protocols to take advantage of the many new developments and applications that have occurred for cellular and molecular investigations. The authors contribute proven protocols for many variations of PRINS and in situ PCR, including a new, fast multicolor PRINS method for identifying human chromosomes, PRINS protocols for in situ detection of unique sequences and point mutations, a new dideoxy PRINS procedure, and in situ PCR and PRINS protocols for plants. They describe, in detail, various applications of PRINS in human cells (detection of gene deletions in cancer, detection of fetal cells in maternal blood, and assessment of aneuploidy in brain tissues and embryos) as well as in plant cells. Readily reproducible in situ PCR techniques are also presented for the detection of cytomegalovirus and for use in combination with microdissection. In situ RT-PCR techniques are also given for use in plant or cancer investigation. The protocols follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and highly practical, PRINS and In Situ PCR Protocols offers molecular biologists, cytogeneticists, and cytologists a cutting-edge guide to today's new in situ labeling technology.