Molecular Protocols in Transfusion Medicine

ISBN-13: 9780122093708 / Angielski / Miękka / 2000 / 186 str.

Greg Denomme; Gregory A. Denomme; Gregory A. Donomme

Opis

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)

Spis treści

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

3.13 Single-strand Conformation Polymorphism (SSCP)

3.14 Optimizing a PCR

3.15 Gene Zygosity Assessment by Quantitative PCR Amplification

3.16 VNTR and STR Analysis by PCR Amplification

3.17 Multiplex PCR Analysis

3.18 Southern Analysis

3.19 References

Part 4: Red Cell Blood Groups

4.1 Terminology

4.2 Clinical Applications

4.3 Situations where Genotype and Phenotype do not Agree

4.4 References

4.5 Facts Sheets, Gene Maps and Molecular Protocols

Part 5: Platelet Blood Groups

5.1 Terminology

5.2 Clinical Applications

5.3 References

5.4 Facts Sheets, Gene Maps and Molecular Protocols

Part 6: Neutrophil Blood Groups

6.1 Terminology

6.2 Clinical Applications

6.3 References

6.4 Facts Sheets, Gene Maps and Molecular Protocols

Part 7: Appendices

7.1 Database Information

7.2 Glossary

7.3 References

7.4 Works Sheets and Request Forms

Index

Useful Tables