Title Hemostasis and thrombosis protocols / edited by David J. Perry and K. John Pasi

Published Totowa, N.J. : Humana Press, ©1999

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Description 1 online resource (xiii, 368 pages) : illustrations Series Methods in molecular medicine ; 31 Methods in molecular medicine ; 31. 1543-1894 Contents PART I. INTRODUCTION -- 1. Hemostasis: Components and Processes / K. John Pasi PART II. BASIC TECHNIQUES -- 2. Isolation of DNA and RNA / David J. Perry -- 3. Amplification of DNA and RNA by PCR / David J. Perry -- 4. Direct Sequencing of PCR Products / David J. Perry -- 5. Solid-Phase Sequencing of Biotinylated PCR Products with Streptavidin-Coated Magnetic Beads / David J. Perry -- 6. Automated DNA Sequencing / Helen L. Devereux -- 7. Detection of DNA by Silver Staining / David J. Perry and Flora Peyvandi -- 8. Promoter Studies in Hemostasis / Peter R. Winship and Jonathan R.K. Spray PART III. METHODS OF MUTATIONAL ANALYSIS -- 9. Detection of Mutations and Polymorphisms in Clotting Factors by Denaturing Gradient Gel Electrophoresis / Rainer Schwaab and Winfried Schmidt -- 10. Screening for Mutations in DNA by Single-Stranded Conformation Polymorphism (SSCP) Analysis / David J. Perry -- 11. Screening for DNA Heteroduplexes in the Factor VII Gene Using Ethylene Glycol Gel Electrophoresis of Solvent-Treated ³²P-Labeled PCR Products / Peter Baker -- 12. Detection of Mutations Causing Hemophilia A Using an In Vitro Coupled Transcription and Translation System / Chike Ononye -- 13. Screening for Mutations in the Human Antithrombin Gene by Hydrolink D-5000 and MDE Gel Electrophoresis / David J. Perry PART IV. METHODS FOR ANALYZING INHERITED/ACQUIRED DISORDERS OF HEMOSTASIS -- 14. Detection of Mutations In Hemophilia A Patients by Chemical Cleavage of Mismatch Method / Naushin H. Waseem, Richard Bagnall, Peter M. Green, and Francesco Giannelli -- 15. Inversion Mutation Analysis in Hemophilia A by Restriction Enzyme Analysis and Southern Blotting / Chike Ononye -- 16. Hemophilia B Mutational Analysis / Peter M. Green -- 17. Screening for Candidate Mutations Causing von Willebrand's Disease (vWD) / P. Vince Jenkins -- 18. Use of Intron 40 VNTR I in vWD Gene Tracking / Mohammed S. Enayat and G.J. Surdhar -- 19. Multimeric Analysis of von Willebrand Factor (vWF) / Mohammed S. Enayat -- 20. Identification of Mutations in the Human Factor VII Gene / Peter M. Baker -- 21. Molecular Analysis in Factor XI Deficiency / Karen M. Johnson and John H. McVey -- 22. Mutational Analysis in Antithrombin Deficiency / David J. Perry -- 23. Ectopic Transcript Analysis in Human Antithrombin Deficiency / David J. Perry -- 24. Mutational Analysis of the Human Protein C Gene / Roger Luddington -- 25. Analysis of the Protein S Gene in Protein S Deficiency / Núria Sala and Yolanda Espinosa-Parrilla -- 26. Screening for the G to A Transition at Position 20210 in the 3ʹ-Untranslated Region (UTR) of the Prothrombin Gene / Karen P. Brown -- 27. Screening for the Factor V Leiden Mutation / Karen P. Brown -- 28. Multiplex PCR for Detection of the Prothrombin 3ʹ UTR (C20210A) Polymorphism and the Factor V Leiden Mutation / Gill Mellars, P. Vince Jenkins, and David J. Perry -- 29. Isoelectric Focusing and Immunodetection of Plasma Antithrombin / Martina Daly -- 30. Characterization of Heparin Binding Variants of Antithrombin by Crossed Immunoelectrophoresis in the Presence of Heparin / Martina Daly -- 31. The Determination of Amino Acid Sequence Abnormalities in Proteins by HPLC Peptide Analysis / David Williamson PART V. PLATELET AND MEGAKARYOCYTE ANALYSIS -- 32. Molecular Biological Identification and Characterization of Inherited Platelet Receptor Disorders / Ramesh B. Basani, Mark Richberg, and Mortimer Poncz -- 33. In Vitro Expansion of Megakaryocytes from Peripheral Blood Hematopoietic Progenitors / Michael A. Thornton and Mortimer Poncz -- 34. Molecular Biology Studies with Primary Megakaryocytes / Yaping Shou and Mortimer Poncz Summary In Hemostasis and Thrombosis Protocols, leading experimentalists describe in detail their proven, cutting-edge methods for research on hemostasis and thrombosis, as well as their diagnostics. The techniques range from widely used basic assays to methods that are specialized for mutational analysis and specific disorders. Readily reproducible, these powerful methods can be used to screen for such inherited disorders as hemophilia A and B, von Willebrands Disease, Factor XI and antithrombin deficiency, protein S deficiency, factor V Leiden mutation, and other problems. The protocols reflect the many major advances that have occurred in our understanding of the molecular genetics of normal hemostasis and it variants, as well as of the coagulation cascade itself. Suitable for both diagnostic and research laboratories, Hemostasis and Thrombosis Protocols enables experienced and novice investigators alike readily to master the complexities of research on molecular hemostasis and thrombosis, as well as work toward the development of productive new molecular diagnostics Bibliography Includes bibliographical references and index Notes Master and use copy. Digital master created according to Benchmark for Faithful Digital Reproductions of Monographs and Serials, Version 1. Digital Library Federation, December 2002. http://purl.oclc.org/DLF/benchrepro0212 MiAaHDL English Print version record and online resource digitized 2010 HathiTrust Digital Library committed to preserve pda MiAaHDL In Springer Protocols Subject Blood coagulation disorders -- Molecular aspects -- Laboratory manuals Blood -- Coagulation -- Laboratory manuals Thrombosis -- Laboratory manuals Blood Coagulation Disorders -- genetics Hemostasis -- genetics DNA Mutational Analysis Sequence Analysis, DNA Thrombosis -- genetics MEDICAL -- Hematology. Blood coagulation disorders Blood -- Diseases -- Molecular aspects Blood -- Coagulation Thrombosis Bloedstolling. Trombose. Wetenschappelijke technieken. Genre/Form Laboratory manuals Laboratory manuals. Manuels de laboratoire. Form Electronic book Author Perry, David (David J.) Pasi, K. John. ISBN 9781592592487 1592592481 9780585244013 0585244014 1280830468 9781280830464

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