Hemostasis -- Basic Techniques -- Isolation of DNA and RNA -- Amplification of DNA and RNA by PCR -- Direct Sequencing of PCR Products -- Solid-Phase Sequencing of Biotinylated PCR Products with Streptavidin-Coated Magnetic Beads -- Automated DNA Sequencing -- Detection of DNA by Silver Staining -- Promoter Studies in Hemostasis -- Methods of Mutational Analysis -- Detection of Mutations and Polymorphisms in Clotting Factors by Denaturing Gradient Gel Electrophoresis -- Screening for Mutations in DNA by Single-Stranded Conformation Polymorphism (SSCP) Analysis -- Screening for DNA Heteroduplexes in the Factor VII Gene Using Ethylene Glycol Gel Electrophoresis of Solvent-Treated 32P-Labeled PCR Products -- Detection of Mutations Causing Hemophilia A Using an In Vitro Coupled Transcription and Translation System -- Screening for Mutations in the Human Antithrombin Gene by Hydrolink D-5000™ and MDE™ Gel Electrophoresis -- Methods for Analyzing Inherited/Acquired Disorders of Hemostasis -- Detection of Mutations in Hemophilia A Patients by Chemical Cleavage of Mismatch Method -- Inversion Mutation Analysis in Hemophilia A by Restriction Enzyme Analysis and Southern Blotting -- Hemophilia B Mutational Analysis -- Screening for Candidate Mutations Causing von Willebrand's Disease (vWD) -- Use of Intron 40 VNTR I in vWD Gene Tracking -- Multimeric Analysis of von Willebrand Factor -- Identification of Mutations in the Human Factor VII Gene -- Molecular Analysis in Factor XI Deficiency -- Mutational Analysis in Antithrombin Deficiency -- Ectopic Transcript Analysis in Human Antithrombin Deficiency -- Mutational Analysis of the Human Protein C Gene -- Analysis of the Protein S Gene in Protein S Deficiency -- Screening for the G to A Transition at Position 20210 in the 3?-Untranslated Region (UTR) of the Prothrombin Gene -- Screening for the Factor V Leiden Mutation -- Multiplex PCR for Detection of the Prothrombin 3?-UTR (G20210A) Polymorphism and the Factor V Leiden Mutation -- Isoelectric Focusing and Immunodetection of Plasma Antithrombin -- Characterization of Heparin Binding Variants of Antithrombin by Crossed Immunoelectrophoresis in the Presence of Heparin -- The Determination of Amino Acid Sequence Abnormalities in Proteins by HPLC Peptide Analysis -- Platelet and Megakaryocyte Analysis -- Molecular Biological Identification and Characterization of Inherited Platelet Receptor Disorders -- In Vitro Expansion of Megakaryocytes from Peripheral Blood Hematopoietic Progenitors -- Molecular Biology Studies with Primary Megakaryocytes.

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.