Molecular Toxicology Protocols / Edition 1

Molecular Toxicology Protocols / Edition 1

by Phouthone Keohavong
     
 

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ISBN-10: 1617373478

ISBN-13: 9781617373473

Pub. Date: 04/08/2014

Publisher: Springer-Verlag New York, LLC

Keohavong and Grant (environmental and occupational health, U. of Pittsburgh) introduce this volume as one addressed to everyone interested in applying molecular methods to clinical populations, yet concede that there is no consensus on the meaning of "molecular." Nevertheless, contributors from universities and cancer centers worldwide describe state-of-the-art

Overview

Keohavong and Grant (environmental and occupational health, U. of Pittsburgh) introduce this volume as one addressed to everyone interested in applying molecular methods to clinical populations, yet concede that there is no consensus on the meaning of "molecular." Nevertheless, contributors from universities and cancer centers worldwide describe state-of-the-art molecular techniques from the field of genetic toxicology to detect genotoxic exposures, identify the agent(s) involved, and treat exposed populations. Emphasizing studies of the exposed individual rather than the agent, 38 chapters provide detailed instructions for laboratory methods for detecting, assessing, and characterizing DNA damage and repair. Annotation ©2004 Book News, Inc., Portland, OR

Product Details

ISBN-13:
9781617373473
Publisher:
Springer-Verlag New York, LLC
Publication date:
04/08/2014
Series:
Methods in Molecular Biology Series, #291
Edition description:
Softcover reprint of hardcover 1st ed. 2005
Pages:
489
Product dimensions:
6.10(w) x 9.25(h) x 0.04(d)

Table of Contents

Prefacev
Contributorsxi
Part IAnalysis of DNA Adducts
1[superscript 32]P-Postlabeling Analysis of DNA Adducts3
2Modification of the [superscript 32]P-Postlabeling Method to Detect a Single Adduct Species as a Single Spot13
3DNA Isolation and Sample Preparation for Quantification of Adduct Levels by Accelerator Mass Spectrometry21
4Fluoroimaging-Based Immunoassay of DNA Photoproducts in Ultraviolet-B-Irradiated Tadpoles29
5Analysis of DNA Strand Cleavage at Abasic Sites39
Part IIDetection of Chromosomal and Genome-Wide Damage
6Premature Chromosome Condensation in Human Resting Peripheral Blood Lymphocytes for Chromosome Aberration Analysis Using Specific Whole-Chromosome DNA Hybridization Probes49
7Mutagen-Induced Chromatid Breakage as a Marker of Cancer Risk59
8Flow Cytometric Analysis of Micronuclei in Erythrocytes69
9The Comet Assay: A Sensitive Genotoxicity Test for the Detection of DNA Damage85
10Computerized Image Analysis Software for the Comet Assay97
11The Comet-FISH Technique: A Tool for Detection of Specific DNA Damage and Repair107
12DNA Double-Strand Break Damage and Repair Assessed by Pulsed-Field Gel Electrophoresis121
Part IIIDetection and Characterization of Surrogate Gene Mutation
13Analysis of In Vivo Mutation in the Hprt and Tk Genes of Mouse Lymphocytes133
14Quantifying In Vivo Somatic Mutations Using Transgenic Mouse Model Systems145
15Methods for Detecting Somatic Mutations In Vitro: The Human T-Cell Cloning Assay Selecting for HPRT Mutants155
16Molecular Analysis of Mutations in the Human HPRT Gene161
17Simultaneous Quantification of t(14;18) and HPRT Exon 2/3 Deletions in Human Lymphocytes171
18The GPA In Vivo Somatic Mutation Assay179
19Flow Cytometric Measurement of Mutant T Cells With Altered Expression of TCR: Detecting Somatic Mutations in Humans and Mice197
Part IVDetection and Characterization of Cancer Gene Mutation
20Mutation Screening of the TP53 Gene by Temporal Temperature Gradient Gel Electrophoresis207
21Analysis of K-RAS and P53 Mutations in Sputum Samples217
22Allele-Specific Competitive Blocker-PCR Detection of Rare Base Substitution235
23Gel-Based Nonradioactive Single-Strand Conformational Polymorphism and Mutation Detection: Limitations and Solutions247
24Detection and Characterization of Oncogene Mutations in Preneoplastic and Early Neoplastic Lesions263
25Detection of DNA Double-Strand Breaks and Chromosome Translocations Using Ligation-Mediated PCR and Inverse PCR279
Part VAnalysis of DNA Repair Mechanisms
26Microsatellite Instability: An Indirect Assay to Detect Defects in the Cellular Mismatch Repair Machinery293
27Unscheduled DNA Synthesis: A Functional Assay for Global Genomic Nucleotide Excision Repair303
28Analysis of DNA Repair Using Transfection-Based Host Cell Reactivation321
29An Immunoassay for Measuring Repair of Ultraviolet Photoproducts337
30Analysis of DNA Double-Strand Break Repair by Nonhomologous End Joining in Cell-Free Extracts From Mammalian Cells351
31Measuring Recombination Proficiency in Mouse Embryonic Stem Cells373
Part VIArray Technologies
32Strategies for Measurement of Biotransformation Enzyme Gene Expression387
33Genotyping Technologies: Application to Biotransformation Enzyme Genetic Polymorphism Screening399
34TaqMan Fluorogenic Detection System to Analyze Gene Transcription in Autopsy Material415
35Development of Quantitative Reverse Transcriptase PCR Assays for Measuring Gene Expression423
Part VIIApoptosis
36Quantification of Selective Phosphatidylserine Oxidation During Apoptosis449
37Quantitative Method of Measuring Phosphatidylserine Externalization During Apoptosis Using Electron Paramagnetic Resonance Spectroscopy and Annexin-Conjugated Iron457
38Detection of Programmed Cell Death in Cells Exposed to Genotoxic Agents Using a Caspase Activation Assay465
Index473

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