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Molecular Protocols in Transfusion Medicine
1st Edition - April 27, 2000
Authors: Gregory A. Denomme, Maria Rios, Marion E. Reid
Language: English
Paperback ISBN:9780122093708
9 7 8 - 0 - 1 2 - 2 0 9 3 7 0 - 8
eBook ISBN:9780080536903
9 7 8 - 0 - 0 8 - 0 5 3 6 9 0 - 3
This essential methods manual for immunohematologists (or hematologists and immunohematologists) provides information on genes that encode antigens on red blood cells, platelets…Read more
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This essential methods manual for immunohematologists (or hematologists and immunohematologists) provides information on genes that encode antigens on red blood cells, platelets and neutrophils. The book begins by covering general concepts in molecular biology and specific protocols such as DNA preparation, PCR-RFLP and allele-specific PCR. Information on the erythrocyte, platelet and neutrophil antigen systems and the molecular basis of polymorphisms are presented clearly in a gene facts sheet format. Database accession numbers and useful adjuncts such as Request forms, worksheets for PCR/enzyme digests also serve to benefit the user. The information is clearly presented and easily accessible and is complemented by the excellent diagrams and tabular material. This book is invaluable for both new and experienced researchers in the field and other related disciplines.
Essential for hematologists and those involoved in tissue typing and the study of human genetic polymorphisms
Presents clearly and concisely the information on a particular variant and the technique used to detect it
Organized by antigen and provides sequences of polymorphisms and primers
Details the general concepts and critical information on genes, their products, and sources of relevant nucleic acids
Includes protocols that allow investigators to set up assays with minimal effort (protocols include primers, reagents, reaction conditions, sizes of amplified products, restriction fragment digests, and the relevant safety information)
Provides information that helps interpret results in clinical settings
Contains additional sources of information (e.g., key references, web site addresses, glossary, Database accession numbers, request forms, and worksheets for PCR/enzyme digests)
Preface
List of Abbreviations
Part 1: Introduction
1.1 Background
1.2 Aim, Scope and Purpose of the Book
1.3 Organization of the Book
1.4 Terminology
1.5 Regulatory Compliance
1.6 References
Part 2: General Considerations
2.1 Clinical Information
2.2 Basic Principles of Molecular Analysis
2.3 Troubleshooting
2.4 References
Part 3: Protocols
3.1 Sample Shipment and Storage
3.2 Reagents
3.3 Harvesting WBCs by RBC Sedimentation
3.4 Short-term Cell Cultures from Amniotic Fluid
3.5 Cultured Amniocyte Cell Harvesting
3.6 Cell Preparation and DNA Extraction
3.7 Assessment of DNA Quality, Quantity and Purity
3.8 PCR Amplification
3.9 Restriction Enzyme Digestion for PCR-RFLP Analysis
3.10 Allele-specific PCR
3.11 Agarose Gel Preparation and Electrophoresis
3.12 Acrylamide Gel Preparation and Electrophoresis