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In-Situ PCR Techniques
 
 
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In-Situ PCR Techniques [Plastic Comb]

Omar Bagasra (Author), John Hansen (Author)

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Book Description

June 1997 0471159468 978-0471159469
This book describes comprehensive step-by-step protocols for the delineation of genetic amplification and histological detection techniques. Each procedure has been tested and validated for its sensitivity, precision, and reproducibility, and the authors give advice on the design of primers for PCR applications and on optimizing these protocols for use with plant, insect, and prokaryotic cells.

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Editorial Reviews

From the Publisher

This book describes comprehensive step-by-step protocols for the delineation of genetic amplification and histological detection techniques. Each procedure has been tested and validated for its sensitivity, precision, and reproducibility, and the authors give advice on the design of primers for PCR applications and on optimizing these protocols for use with plant, insect, and prokaryotic cells.

From the Back Cover

In situ gene amplification techniques offer tremendous potential as aids to clinical diagnosis through their ability to detect a single copy of a specific microbial, neoplastic, messenger, or mutated nucleic acid sequence in a cell smear, cell suspension, tissue section, or chromosome. In situ hybridization, applied in combination with the polymerase chain reaction (PCR), can be expected to fuel accelerated developments in the understanding of embryogenesis, organogenesis, and cell differentiation, as well as the pathogenesis of numerous disease processes. But the procedures are cumbersome and fraught with potential variables, and experimental results are difficult to reproduce.

In Situ PCR Techniques addresses this problem directly, with comprehensive step-by-step protocols for the delineation of genetic amplification and histological detection techniques. Each procedure has been tested and validated for its sensitivity, precision, and reproducibility, and the authors give advice on the design of primers for PCR applications and on optimizing these protocols for use with plant, insect, and prokaryotic cells. They facilitate the repetition of published experiments by providing the kinds of hints, tips, and laboratory secrets that are often left out of scientific papers. They also demonstrate clearly and thoroughly the efficacy of in situ PCR as an alternative to standard and fluorescence in situ hybridization for detecting a single copy of a mutant gene, a virus, or a very low-abundance message in individual cells, while preserving the morphology of the cell and the tissue in which it resides.

Topics covered include:
* A review of the in situ PCR technique
* Step-by-step protocols for in situ PCR techniques
* Optimization of annealing temperatures for specific primers
* Preparation of glass slides and tissues
* Selection of DNA and RNA targets

Supplemented with numerous helpful graphics and illustrations, In Situ PCR Techniques is immensely useful, not merely to the novice, but also to experienced researchers in investigative pathology, virology, and gene therapy, as well as in developmental biology, immunology, plant molecular biology, entomology, and the neurosciences.

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Inside This Book (learn more)
First Sentence:
This chapter is a primer for those investigators for whom molecular techniques are a relatively new experience. Read the first page
Key Phrases - Statistically Improbable Phrases (SIPs): (learn more)
amplification cocktail, situ amplification, gene cones, situ polymerase chain reaction, cycling regime, oligo probes, thermal cycler, situ reactions, incubate the slides, nucleic acid targets, amplification protocol, microvascular endothelial cells, amplification procedure, original locus
Key Phrases - Capitalized Phrases (CAPs): (learn more)
Perkin Elmer, Repeat Steps, World Wide Web
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