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Diagnostic Molecular Biology
- 2nd Edition - June 29, 2023
- Author: Chang-Hui Shen
- Language: English
- Paperback ISBN:9 7 8 - 0 - 3 2 3 - 9 1 7 8 8 - 9
- eBook ISBN:9 7 8 - 0 - 3 2 3 - 9 8 6 0 9 - 0
Diagnostic Molecular Biology, Second Edition provides an up-to-date comprehensive overview of the latest advances in molecular diagnostics and the fundamental molecular aspects i… Read more
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Request a sales quoteDiagnostic Molecular Biology, Second Edition provides an up-to-date comprehensive overview of the latest advances in molecular diagnostics and the fundamental molecular aspects in this area. The book delves into the essential principles behind the transmission and expression of genetic information, including nucleic acids, proteins, and chromosomes, specifically their genome structure, life cycle, and evolution. The latest methods and instrumentation in the field are examined with detailed procedures and explanations provided for each technique. A range of techniques are also investigated, such as those used in sequencing analysis, genome and transcriptome analysis, and nucleic acid amplitification techniques. This book covers the collection, isolation, and purification of DNA, RNA, and protein from different sources. Additionally, discussion on the applications of next-generation sequencing (NGS) and the role of NGS in the future of precision medicine is included. The book also examines the use of molecular diagnostics for chromosomal disorders, inherited diseases, and infectious diseases. Moreover, this new edition features recent advances in the detection of respiratory virus infections in humans, including that of SARS-CoV2. Diagnostic Molecular Biology, Second Edition, is an ideal reference for researchers, lab technicians, and students in medical laboratory science, molecular biology, microbiology, virology, and related fields.
- Explores techniques used for diagnosis at the molecular level
- Explains the molecular mechanism of genetic disorders and infectious diseases
- Enhances understanding of the fundamentals of proteins, nucleic acids and cellular activities based on proteins and/or nucleic acids
- Includes clinical applications of molecular diagnostics for a variety of disorders and diseases
researchers and technicians in the lab, pathologists, and students in molecular biology, microbiology and virology clinical laboratory scientists and technicians
- Cover image
- Title page
- Table of Contents
- Copyright
- Dedication
- Preface
- Acknowledgement
- Chapter 1. Nucleic acids: DNA and RNA
- DNA/RNA is genetic material
- The components of nucleic acids
- The structure of DNA molecules
- Conformational changes of the DNA double helix
- Alternative forms of DNA structure
- Physical properties of nucleic acids
- Alternative forms of RNA molecules and their secondary structure
- RNA folding
- Chapter 2. Nucleic acid-based cellular activities – DNA replication, damage, and repair
- DNA replication
- The process of DNA replication
- Proteins required for DNA replication in E. coli
- Coupling DNA replication
- Replication patterns
- Eukaryotic DNA replication
- Fidelity of DNA replication
- DNA mutation and repair
- DNA damage
- Damage repair: single-strand damage repair
- Damage repair: double-strand break repair
- Chapter 3. Gene expression: transcription of the genetic code
- General features of RNA synthesis
- Promoters
- RNA polymerase in Escherichia coli
- The process of transcription
- The regulation of prokaryotic transcription
- Transcription in eukaryotes
- The order of events in eukaryotic transcription initiation
- Transcription elongation through the nucleosomal barrier
- Maturation process of eukaryotic mRNA
- Noncoding RNAs
- Chapter 4. Gene expression: translation of the genetic code
- The structure and properties of amino acids
- Classifications of amino acids
- Protein structure
- Folding into three-dimensional protein structure
- The genetic code
- Translational machinery: ribosome
- Translational machinery: tRNA
- Preparation for the translation
- Aminoacyl-tRNA synthetases
- Prokaryotic translation
- Eukaryotic translation
- Protein folding
- Posttranslational modification of proteins
- Chapter 5. The genome
- The genome in bacteria
- The genome in viruses
- Eukaryotic chromosome structure
- The eukaryotic cell cycle and chromosome dynamics
- Higher order chromatin structure
- Unique animal chromosomes
- Specialized DNA sequences in eukaryotic chromosomes
- Complexity and gene number
- The features of genomic sequences
- Organelle genomes
- Genome evolution
- Mechanisms of genome evolution
- Evolution of the prokaryotic genome
- Chapter 6. Extraction and purification of nucleic acids
- The importance of DNA extraction
- Genomic DNA extraction from whole blood sample
- DNA extraction from dry blood spots
- Noninvasive human DNA isolation
- DNA preparation from microorganisms
- Conventional methods for nucleic acid purification
- Solid-phase nucleic acid extraction
- Magnetic beads extraction
- Microfluidic-based nucleic acid extraction
- DNA preparation from formalin-fixed, paraffin-embedded tissues
- RNA preparation
- Extraction of RNA
- Chapter 7. Quantification and analysis of nucleic acids
- Determining DNA purity and yield
- Molecular separation of DNA
- Agarose gel electrophoresis
- Polyacrylamide gels
- Special application of polyacrylamide gel electrophoresis
- Pulsed-field gel electrophoresis for large DNA molecules
- Capillary electrophoresis
- Different modes of CE
- Micro/nanofluidic devices for DNA/RNA separation
- Chapter 8. Extraction and purification of proteins
- Protein extract preparation
- Cell lysis conditions
- Preparation of clean protein samples
- Protein extraction from archival formalin-fixed paraffin-embedded tissues
- Protein purification through column chromatography
- High performance liquid chromatography
- Chapter 9. Quantification and analysis of proteins
- UV spectroscopy assay
- Colorimetric protein assay techniques
- Fluorescence-based assays
- The choice of measurement format
- Molecular separation of protein through electrophoresis
- Gel-based proteomics: protein separation by two-dimensional gel electrophoresis
- Sample preparation and sample buffer constituents for 2DE
- Gel free protein analysis—capillary electrophoresis
- Protein analysis and characterization with mass spectrometry (MS)
- Chapter 10. Amplification of nucleic acids
- Conventional amplification technique: the polymerase chain reaction (PCR)
- Parameters that affect PCR
- Modified nucleic acid amplification techniques
- Quantitative PCR
- Double-stranded DNA binding dyes for qPCR
- Fluorophore-labeled oligonucleotides for qPCR
- Isothermal amplification techniques
- Chapter 11. Characterization of nucleic acids and proteins
- Nucleases
- Restriction enzyme digestion frequency and analysis
- Polymerases
- DNA modifying enzymes and ligases
- The application of nucleic acid hybridization
- Detection methods for nucleic acid hybridization
- Array-based hybridization
- DNA microarray
- Protein immunodetection technique—Western blot
- Protein immunodetection technique—enzyme-linked immunosorbent assay (ELISA)
- Protein microarrays
- Chapter 12. Techniques in sequencing
- Chain-termination DNA sequencing technique
- Chain-termination sequencing-based genome sequencing
- Assembly approach for clone contigs—clone fingerprinting techniques
- Next-generation sequencing (NGS)
- The third-generation sequencer
- NGS data analysis
- The applications of NGS technologies
- Chapter 13. Genome and transcriptome analysis
- Genome-Wide sampling sequencing for genetic polymorphisms
- The association of the genetic polymorphisms with human diseases
- Metabolomics for GWAS: linking the genome to the metabolome
- Direct sequencing of total RNA—transcriptome sequencing
- Transcriptome sequencing: whole transcriptome shotgun sequencing—EST (expressed sequence tag) sequencing procedure
- Transcriptome sequencing: whole transcriptome shotgun sequencing—RNA-seq procedure
- Whole transcriptome tag/target sequencing with restriction digestion—SAGE (serial analysis of gene expression)
- Whole transcriptome tag/target sequencing without restriction digestion
- Metagenomics
- Chapter 14. Molecular diagnosis of chromosomal disorders
- The features of centromere
- Conventional cytogenetics
- Abnormalities of chromosome number
- Abnormalities of chromosome structure
- Molecular cytogenetics: fluorescence in situ hybridization (FISH)
- FISH probes
- Advances in FISH technology
- FISH applications in clinical diagnostics
- Molecular cytogenetics: comparative genomic hybridization
- Applications of CGH analysis
- Chromosomal disorders: numerical chromosome abnormalities
- Chromosomal disorders: sex chromosome abnormalities
- Chromosomal disorders: structural chromosome abnormalities
- The causes of chromosome abnormalities
- Chapter 15. Molecular diagnosis of gene mutation and inherited diseases
- Variations that cause human disease
- Choice of techniques to detect genome alteration that causes disease
- Single-gene disorders
- Polygenic disorders and gene clustering
- Mitochondria disorders
- Chapter 16. Molecular diagnosis of infectious diseases
- Detection of bacterial pathogens, fungal pathogens, and viruses
- Detection of bacteria
- Molecular detection and characterization of fungal pathogens
- Molecular detection of viruses
- Emerging technology: MALDI-TOF MS for microbial species identification
- Chapter 17. The biology of SARS-CoV-2 and the molecular diagnosis of COVID-19
- Biology of coronaviruses
- SARS-CoV-2 proteins and their functions
- SARS-CoV-2 life cycle
- SARS-CoV-2 variants
- COVID-19 signs and symptoms
- Common symptoms of COVID-19
- Molecular diagnosis of COVID-19
- RT-PCR
- RT-LAMP
- CRISPR-Cas13-based diagnosis—SHERLOCK
- Immunological detection of COVID-19
- Vaccine
- RNA COVID-19 vaccines
- Treatment
- Chapter 18. Guidance for molecular diagnostic laboratory
- The need for molecular analysis
- Preanalytical standards
- Choice of analytical method
- Analytical phase
- Storage of DNA and RNA
- Postanalytical analysis
- General administrative guidelines
- Accreditation based on international standard
- Regulation of molecular diagnosis in the US
- Challenges for molecular diagnosis in a clinical laboratory
- Bibliography
- Index
- No. of pages: 588
- Language: English
- Edition: 2
- Published: June 29, 2023
- Imprint: Academic Press
- Paperback ISBN: 9780323917889
- eBook ISBN: 9780323986090
CS
Chang-Hui Shen
Chang-Hui Shen, PhD, is a Professor of Biology and Biochemistry at the College of Staten Island and the Graduate Center, City University of New York. He earned his B.S. and M.S. at the National Chung-Hsing University, Taiwan, and his Ph.D. at the University of Edinburgh, UK. He was a visiting fellow of the National Institutes of Health before joining the City University of New York.
Dr. Shen’s research focuses on the mechanism of gene expression, with an emphasis on epigenetic regulation in gene activation. His lab utilizes the breadth of current and advanced techniques in molecular biology and biochemistry. He has published dozens of articles throughout his career, and his research has been supported by several agencies, including NATO, NIH, and NSF.
Affiliations and expertise
Professor of Biology and Chair of the Biology Department at the College of Staten Island, City University of New York, New York, USARead Diagnostic Molecular Biology on ScienceDirect