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Molecular Diagnostics: Fundamentals, Methods, and Clinical Applications 3rd Edition by Lela Buckingham, ISBN-13: 978-0803668294

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Molecular Diagnostics: Fundamentals, Methods, and Clinical Applications 3rd Edition by Lela Buckingham, ISBN-13: 978-0803668294

[PDF eBook eTextbook]

  • Publisher: ? F.A. Davis Company; Third edition (February 22, 2019)
  • Language: ? English
  • ?576 pages
  • ISBN-10: ? 0803668295
  • ISBN-13: ? 978-0803668294

The only book you need to become certified.

Meet the challenges of this rapidly expanding field.

Molecular diagnostics continues to grow in importance in the clinical laboratory. This respected text will prepare your students with a grounding in the fundamental principles of molecular biology. current methods, and their clinical applications.

With a focus on the application of molecular concepts to diagnostic purposes, the text explains and illustrate the use and interpretation of molecular-based assays in patient care. Now with an expanded discussion of nucleic acid sequencing, with added emphasis on next generation sequencing (NGS), and updated coverage of proteomics and mass spectrometry applications, your students will have the most current information available today.

Table of Contents:

Cover
Title Page
Copyright Page
Dedication
Preface
Reviewers
Acknowledgments
Contents
Section I: Fundamentals of Molecular Biology: An Overview
1 Nucleic Acids and Proteins
DNA
DNA STRUCTURE
Nucleotides
Nucleic Acid
DNA REPLICATION
Polymerases
ENZYMES THAT METABOLIZE DNA
Restriction Enzymes
DNA Ligase
Other DNA Metabolizing Enzymes
RECOMBINATION IN SEXUALLY REPRODUCING ORGANISMS
RECOMBINATION IN ASEXUAL REPRODUCTION
Conjugation
Transduction
Transformation
PLASMIDS
RNA
Transcription
Transcription Initiation
Transcription Elongation
Transcription Termination
TYPES/STRUCTURES OF RNA
Ribosomal RNA
Messenger RNA
Small Nuclear RNA
Transfer RNA
Other RNAs
RNA POLYMERASES
OTHER RNA-METABOLIZING ENZYMES
Ribonucleases
RNA Helicases
PROTEINS AND THE GENETIC CODE
Amino Acids
Genes
The Genetic Code
TRANSLATION
Amino Acid Charging
Protein Synthesis
2 Gene Expression and Epigenetics
TRANSCRIPTION
Transcription Initiation
Transcription Elongation
Transcription Termination
REGULATION OF TRANSCRIPTION
Regulation of Messenger RNA Synthesis at Initiation
Post-Transcriptional Regulation
Post-Translational Regulation
EPIGENETICS
Histone Modification
Nucleic Acid Methylation
CLASSIFICATION OF EPIGENETIC FACTORS
NONCODING RNAs
MicroRNAs
Small Interfering RNAs
Other Small RNAs
Long Noncoding RNAs
Section II: Common Techniques in Molecular Biology
3 Nucleic Acid Extraction Methods
ISOLATION OF DNA
Preparing the Sample
DNA Isolation Chemistries
ISOLATION OF RNA
Total RNA
Specimen Collection
RNA Isolation Chemistries
MEASUREMENT OF NUCLEIC ACID QUALITY AND QUANTITY
Electrophoresis
Spectrophotometry
Fluorometry
Microfluidics
4 Resolution and Detection of Nucleic Acids
ELECTROPHORESIS OF NUCLEIC ACIDS
GEL SYSTEMS
Agarose Gels
Polyacrylamide Gels
CAPILLARY ELECTROPHORESIS
BUFFER SYSTEMS
Buffer Additives
ELECTROPHORESIS EQUIPMENT
Gel Loading
DETECTION SYSTEMS
Fluorescent Dyes
Silver Stain
5 Analysis and Characterization of Nucleic Acids and Proteins
RESTRICTION ENZYME MAPPING OF DNA
CRISPR ENZYME SYSTEMS
HYBRIDIZATION TECHNOLOGIES
Southern Blots
PROBE HYBRIDIZATION
Northern Blots
Western Blots
PROBES
DNA Probes
RNA Probes
Other Nucleic Acid Probe Types
Protein Probes
Probe Labeling
Nucleic Acid Probe Design
HYBRIDIZATION CONDITIONS, STRINGENCY
DETECTION SYSTEMS
INTERPRETATION OF RESULTS
ARRAY-BASED HYBRIDIZATION
Dot/Slot Blots
Genomic Array Technology
SOLUTION HYBRIDIZATION
6 Nucleic Acid Amplification
TARGET AMPLIFICATION
Polymerase Chain Reaction
Transcription-Based Amplification Systems
Genomic Amplification Methods
PROBE AMPLIFICATION
Ligase Chain Reaction
Strand Displacement Amplification
Qß Replicase
SIGNAL AMPLIFICATION
Branched DNA Amplification
Hybrid Capture Assays
Cleavage-Based Amplification
Cycling Probe
7 Chromosomal Structure and Chromosomal Mutations
CHROMOSOMAL STRUCTURE AND ANALYSIS
Chromosomal Compaction and Histones
Chromosome Morphology
Visualizing Chromosomes
DETECTION OF GENOME AND CHROMOSOMAL MUTATIONS
Karyotyping
Fluorescence In Situ Hybridization
COMPARATIVE GENOME HYBRIDIZATION (CGH)
8 Gene Mutations
TYPES OF GENE MUTATIONS
DETECTION OF GENE MUTATIONS
Biochemical Methods
Nucleic Acid Analyses
GENE VARIANT NOMENCLATURE
GENE NAMES
9 DNA Sequencing
DIRECT SEQUENCING
Manual Sequencing
Automated Fluorescent Sequencing
PYROSEQUENCING
BISULFITE DNA SEQUENCING
RNA SEQUENCING
NEXT-GENERATION SEQUENCING
Gene Panels
NGS Library Preparation
Targeted Libraries
Sequencing Platforms
Sequence Quality
Filtering and Annotation
BIOINFORMATICS
THE HUMAN GENOME PROJECT
Variant Associations With Phenotype
Section III: Techniques in the Clinical Laboratory
10 DNA Polymorphisms and Human Identification
TYPES OF POLYMORPHISMS
RFLP TYPING
Genetic Mapping With RFLPs
RFLP and Parentage Testing
Human Identification Using RFLPs
STR TYPING BY PCR
STR Analysis
Y-STR
LINKAGE ANALYSIS
BONE MARROW ENGRAFTMENT TESTING USING DNA POLYMORPHISMS
PSTR Testing
Post-Transplant Engraftment Testing
QUALITY ASSURANCE FOR SURGICAL SECTIONS USING STR
SINGLE-NUCLEOTIDE POLYMORPHISMS
The Human Haplotype Mapping (HapMap) Project
MITOCHONDRIAL DNA POLYMORPHISMS
OTHER IDENTIFICATION METHODS
Protein-Based Identification
Epigenetic Profiles
11 Detection and Identification of Microorganisms
SPECIMEN COLLECTION
SAMPLE PREPARATION
QUALITY ASSURANCE
Controls
Quality Control
Selection of Sequence Targets for Detection of Microorganisms
MOLECULAR DETECTION OF MICROORGANISMS
Bacteria
Viruses
Mycology
Parasites
ANTIMICROBIAL AGENTS
Resistance to Antimicrobial Agents
Molecular Detection of Resistance
MOLECULAR EPIDEMIOLOGY
Molecular Strain Typing Methods for Epidemiological Studies
Comparison of Typing Methods
12 Molecular Detection of Inherited Diseases
THE MOLECULAR BASIS OF INHERITED DISEASES
CHROMOSOMAL ABNORMALITIES
PATTERNS OF INHERITANCE IN SINGLE-GENE DISORDERS
MOLECULAR BASIS OF SINGLE-GENE DISORDERS
Lysosomal Storage Diseases
Factor V Leiden
Prothrombin
Methylenetetrahydrofolate Reductase
Hemochromatosis
Cystic Fibrosis
Cytochrome P-450
SINGLE-GENE DISORDERS WITH NONCLASSICAL PATTERNS OF INHERITANCE
Mutations in Mitochondrial Genes
Nucleotide-Repeat Expansion Disorders
Genomic Imprinting
Multifactorial Inheritance
LIMITATIONS OF MOLECULAR TESTING
13 Molecular Oncology
CLASSIFICATION OF NEOPLASMS
MOLECULAR BASIS OF CANCER
ANALYTICAL TARGETS OF MOLECULAR TESTING
GENE AND CHROMOSOMAL MUTATIONS IN SOLID TUMORS
Human Epidermal Growth Factor Receptor 2, HER2/neu/erb-b2 1 (17q21.1)
Epidermal Growth Factor Receptor, EGFR (7p12)
Kirsten Rat Sarcoma Viral Oncogene Homolog, K-ras (12p12); Neuroblastoma ras, N-ras (1p13); and Harvey Rat Sarcoma Viral Oncogene Homolog, H-ras (11p15)
Ewing Sarcoma, EWS (22q12)
Synovial Sarcoma Translocation, Chromosome 18—Synovial Sarcoma Breakpoint 1 and 2, SYT-SSX1, SYT-SSX2 t(X;18) (p11.2;q11.2)
Paired Box-Forkhead in Rhabdomyosarcoma, PAX3-FKHR, PAX7-FKHR, t(1;13), t(2;13)
Tumor Protein 53, TP53 (17p13)
Ataxia Telangiectasia Mutated Gene, ATM (11q22)
Breast Cancer 1 Gene, BRCA1 (17q21), and Breast Cancer 2 Gene, BRCA2 (13q12)
Von Hippel–Lindau Gene, VHL (3p26)
V-myc Avian Myelocytomatosis Viral-Related Oncogene, Neuroblastoma-Derived, MYCN or n-myc (2p24)
V-Ros Avian UR2 Sarcoma Virus Oncogene Homolog 1 (ROS1) Proto-Oncogene (6q22.1) and Rearranged During Transfection (RET) Proto-Oncogene (10q11)
Anaplastic Lymphoma Receptor Tyrosine Kinase (ALK) Proto-Oncogene, 2p23.1
V-Kit Hardy-Zuckerman 4 Feline Sarcoma Viral Oncogene Homolog, KIT, c-KIT (4q12)
Other Molecular Abnormalities
Microsatellite Instability
Loss of Heterozygosity
Liquid Biopsy
MOLECULAR ANALYSIS OF LEUKEMIA AND LYMPHOMA
Gene Rearrangements
Mutations in Hematological Malignancies
Mutation Spectra
14 DNA-Based Tissue Typing
THE MHC LOCUS
HLA POLYMORPHISMS
HLA Nomenclature
MOLECULAR ANALYSIS OF THE MHC
Serological Analysis
DNA-Based Typing
Combining Typing Results
HLA Test Discrepancies
Coordination of HLA Test Methods
ADDITIONAL RECOGNITION FACTORS
Minor Histocompatibility Antigens
Nonconventional MHC Antigens
Killer Cell Immunoglobulin-Like Receptors
MHC DISEASE ASSOCIATION
SUMMARY OF LABORATORY TESTING
15 Quality Assurance and Quality Control in the Molecular Laboratory
SPECIMEN HANDLING
Collection Tubes for Molecular Testing
Precautions
Holding and Storage Requirements
TEST PERFORMANCE
Next-Generation Sequencing
Calibrators and Method Calibration
Controls
QUALITY CONTROL
QUALITY ASSURANCE
INSTRUMENT MAINTENANCE
Instrument Calibration
REAGENTS
Reagent Categories
Chemical Safety
Reagent Storage
Reagent Labeling
PROFICIENCY TESTING
DOCUMENTATION OF TEST RESULTS
Gene Nomenclature
Gene Sequencing Results
REPORTING RESULTS
Appendix A Study Question Answers
Appendix B Answers to Case Studies
Glossary
Index

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