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Hemostasis and Thrombosis Protocols

  • Book
  • © 1999

Overview

Editors:
  1. David J. Perry

    1. Royal Free Hospital, London, UK

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    You can also search for this editor in PubMed   Google Scholar

  2. K. J. Pasi

    1. Royal Free Hospital, London, UK

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

Part of the book series: Methods in Molecular Medicine (MIMM, volume 31)

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Table of contents (34 protocols)

  1. Front Matter

    Pages i-xiii
    Download chapter PDF

  2. Introduction

    1. Front Matter

      Pages 1-1
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    2. Hemostasis

      • K. John Pasi
      Pages 3-21

  3. Basic Techniques

    1. Front Matter

      Pages 23-23
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    2. Isolation of DNA and RNA

      • David J. Perry
      Pages 25-30

    3. Amplification of DNA and RNA by PCR

      • David J. Perry
      Pages 31-38

    4. Direct Sequencing of PCR Products

      • David J. Perry
      Pages 39-48

    5. Solid-Phase Sequencing of Biotinylated PCR Products with Streptavidin-Coated Magnetic Beads

      • David J. Perry
      Pages 49-54

    6. Automated DNA Sequencing

      • Helen L. Devereux
      Pages 55-61

    7. Detection of DNA by Silver Staining

      • David J. Perry, Flora Peyvandi
      Pages 63-64

    8. Promoter Studies in Hemostasis

      • Peter R. Winship, Jonathan R. K. Spray
      Pages 65-81

  4. Methods of Mutational Analysis

    1. Front Matter

      Pages 83-83
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    2. Detection of Mutations and Polymorphisms in Clotting Factors by Denaturing Gradient Gel Electrophoresis

      • Rainer Schwaab, Winfried Schmidt
      Pages 85-104

    3. Screening for Mutations in DNA by Single-Stranded Conformation Polymorphism (SSCP) Analysis

      • David J. Perry
      Pages 105-110

    4. Screening for DNA Heteroduplexes in the Factor VII Gene Using Ethylene Glycol Gel Electrophoresis of Solvent-Treated 32P-Labeled PCR Products

      • Peter Baker
      Pages 111-116

    5. Detection of Mutations Causing Hemophilia A Using an In Vitro Coupled Transcription and Translation System

      • Chike Ononye, P. Vincent Jenkins
      Pages 117-124

    6. Screening for Mutations in the Human Antithrombin Gene by Hydrolink D-5000™ and MDE™ Gel Electrophoresis

      • David J. Perry
      Pages 125-129

  5. Methods for Analyzing Inherited/Acquired Disorders of Hemostasis

    1. Front Matter

      Pages 131-131
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    2. Detection of Mutations in Hemophilia A Patients by Chemical Cleavage of Mismatch Method

      • Naushin H. Waseem, Richard Bagnall, Peter M. Green, Francesco Giannelli
      Pages 133-149

    3. Inversion Mutation Analysis in Hemophilia A by Restriction Enzyme Analysis and Southern Blotting

      • Chike Ononye, P. Vincent Jenkins
      Pages 151-157
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About this book

Laboratory studies in hemostasis have traditionally focused on abn- malities of platelet function or the quantitative and qualitative disorders that affect the proteins involved in blood coagulation. However, over the last 10 years there has been an explosion in our understanding of the molecular bases that underlie many of the inherited and acquired disorders of hemostasis. Many of these disorders are now routinely diagnosed and assessed by methods that involve genotypic analysis. Indeed in the late 1990s the distinction between molecular methods for research and for routine diagnosis is becoming incre- ingly blurred. The techniques and approaches that are used in hemostasis are manifold and published in isolation in a variety of publications. The aim, therefore, of this volume Hemostasis and Thrombosis Protocols is to pull together, into a single volume, the variety of techniques that are frequently used in the field of hemostasis. We have targeted this volume at laboratories who wish to move into the field of molecular hemostasis or who may already have some expe- ence in this area but wish to develop new areas of research and diagnosis. The chapters are wide-ranging and hopefully provide a broad overview of the d- fering applications in which these standard techniques can be used. Though the articles may appear relatively specific, the techniques contained within them are applicable to the study of many different disorders and we hope that they provide a series of ideas and concepts well-suited to problem solving.

Reviews

"...The book by Perry and Pasi confirms ...that with good recipes and kits the DNA analysis of disorders of thrombosis and hemostasis is indeed feasible for many diagnostic laboratories. The book describes in great detail a variety of laboratory protocols aimed primarily at analyzing inherited disorders of the hemostatic system. These protocols vary from simple procedures to detect factor V Leiden and prothrombin 20210 to elaborate techniques for finding mutations in disorders like hemophilia A or protein S deficiency. In addition ...this book is a useful book for those attempting to introduce or maintain DNA analysis in their laboratory...laboratory technicians may find many useful suggestions to improve their work."-Haemostasis

"Each cheater is presented in a recipe format so that the methods can e reproduced. All chapters subdivided into sections for easy reading: introduction, material, methods, and notes. ...The chapters on mutation detection in clotting factor deficiencies will be useful to hemostasis laboratories that are interested in carrier detection, prenatal diagnosis, or research. The chapters on von Willbrand factor multimer analysis, factor V Leiden mutation, an prothrombin G20210A mutation will be useful to hemostasis laboratories that want to set up these important diagnostic tests...This manual does well at meeting its goal of providing a broad overview of techniques used in the analysis of hemostatic disorders and providing strategies for future problem solving. It will be a great utility to hemostasis laboratories that wish to advance their research or diagnostic capabilities."-Modern Pathology

"very valuable for the laboratory staff in charge of gene analysis of known genetic defects in known diseases of hemostasis and thrombosis." - Bloodline

Editors and Affiliations

  • Royal Free Hospital, London, UK

    David J. Perry, K. J. Pasi

Bibliographic Information

  • Book Title: Hemostasis and Thrombosis Protocols

  • Editors: David J. Perry, K. J. Pasi

  • Series Title: Methods in Molecular Medicine

  • DOI: https://doi.org/10.1385/1592592481

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media New York 1999

  • Hardcover ISBN: 978-0-89603-419-8Published: 04 August 1999

  • Softcover ISBN: 978-1-4899-4313-2Published: 09 August 2013

  • eBook ISBN: 978-1-59259-248-7Published: 02 February 2008

  • Series ISSN: 1543-1894

  • Series E-ISSN: 1940-6037

  • Edition Number: 1

  • Number of Pages: XIII, 368

  • Topics: Pathology

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