Hemostasis and thrombosis protocols
Author(s)
Bibliographic Information
Hemostasis and thrombosis protocols
(Methods in molecular medicine / John M. Walker, series editor, 31)
Humana Press, c1999
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Note
Includes bibliographical references and index
Description and Table of Contents
Description
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.
Table of Contents
Part I. Introduction.
Hemostasis: Components and Processes
K. John Pasi
Part II. Basic Techniques.
Isolation of DNA and RNA
David J. Perry
Amplification of DNA and RNA by PCR
David J. Perry
Direct Sequencing of PCR Products
David J. Perry
Solid-Phase Sequencing of Biotinylated PCR Products with Streptavidin-Coated Magnetic Beads
David J. Perry
Automated DNA Sequencing
Helen L. Devereux
Detection of DNA by Silver Staining
David J. Perry and Flora Peyvandi
Promoter Studies in Hemostasis
Peter R. Winship and Jonathan R. K. Spray
Part III. Methods of Mutational Analysis.
Detection of Mutations and Polymorphisms in Clotting Factors by Denaturing Gradient Gel Electrophoresis
Rainer Schwaab and Winfried Schmidt
Screening for Mutations in DNA by Single-Stranded Conformation Polymorphism (SSCP) Analysis
David J. Perry
Screening for DNA Heteroduplexes in the Factor VII Gene Using Ethylene Glycol Gel Electrophoresis of Solvent-Treated 32P-Labeled PCR Products
Peter M. Baker
Detection of Mutations Causing Hemophilia A Using an In Vitro Coupled Transcription and Translation System
Chike Ononye and P. Vincent Jenkins
Screening for Mutations in the Human Antithrombin Gene by Hydrolink D-5000 (TM) and MDE (TM) Gel Electrophoresis
David J. Perry
Part IV. Methods for Analyzing Inherited/Acquired Disorders of Hemostasis
Detection of Mutations In Hemophilia A Patients by Chemical Cleavage of Mismatch Method
Naushin H. Waseem, Richard Bagnall, Peter M. Green, and Francesco Giannelli
Inversion Mutation Analysis in Hemophilia A by Restriction Enzyme Analysis and Southern Blotting
Chike Ononye and P. Vincent
Hemophilia B Mutational Analysis
Peter M. Green
Screening for Candidate Mutations Causingvon Willebrand's Disease (vWD)
P. Vincent Jenkins
Use of Intron 40 VNTR I in vWD Gene Tracking
Mohammed S. Enayat and Gurcharan J. Surdhar
Multimeric Analysis of von Willebrand Factor (vWF)
Mohammed S. Enayat
Identification of Mutations in the Human Factor VII Gene
Peter M. Baker
Molecular Analysis in Factor XI Deficiency
Karen M. Johnson and John H. McVey
Mutational Analysis in Antithrombin Deficiency
David J. Perry
Ectopic Transcript Analysis in Human Antithrombin Deficiency
David J. Perry
Mutational Analysis of the Human Protein C Gene
Roger Luddington
Analysis of the Protein S Gene in Protein S Deficiency
Nuria Sala and Yolanda Espinosa-Parrilla
Screening for the G to A Transition at Position 20210 in the 3'-Untranslated Region (UTR) of the Prothrombin Gene
Karen P. Brown
Screening for the Factor V Leiden Mutation
Karen P. Brown
Multiplex PCR for Detection of the Prothrombin 3' UTR (C20210A) Polymorphism and the Factor V Leiden Mutation
Gillian Mellars, P. Vincent Jenkins, and David J. Perry
Isoelectric Focusing and Immunodetection of Plasma Antithrombin
Martina Daly
Characterization of Heparin Binding Variants of Antithrombin by Crossed Immunoelectrophoresis in the Presence of Heparin
Martina Daly
The Determination of Amino Acid Sequence Abnormalities in Proteins by HPLC Peptide Analysis
David Williamson
Part V. Platelet and Megakaryocyte Analysis
Molecular Biological Identification and Characterization of Inherited Platelet Receptor Disorders
Ramesh B. Basani, Mark Richberg, and Mortimer Poncz
In Vitro Expansion of Megakaryocytes from Peripheral Blood Hematopoietic Progenitors
Michael A. Thornton and Mortimer Poncz
Molecular Biology Studies with Primary Megakaryocytes
Yaping
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