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Trinucleotide Repeat Protocols : Methods in Molecular Biology - Yoshinori Kohwi

Trinucleotide Repeat Protocols

Methods in Molecular Biology

By: Yoshinori Kohwi (Editor)


Published: 18th June 2004
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Established leaders in trinucleotide repeat disease describe in step-by-step detail their best techniques for studying trinucleotide pathology at the molecular level. The protocols cover a variety of targets, ranging from DNA and RNA to proteins and whole animals, and focus not only on causal genes, but also on their consequent affected products, such as transcription factors, neurotransmitter receptors, proteasomes, and mitochondria/oxidation damage. Experimental systems employed include E. coli, yeast, C. elegans, mouse, and generally take a clinical point of view. The authors utilize a wide range of techniques, including gel electrophoresis, quantitative RT-PCR, immunological analysis, antibody usage and its applications, receptor assays using radioisotope handling, gene delivery by virus, brain cell and organotypic cultures, gender dependency, and neuron structure analysis.

Prefacep. v
Contributorsp. ix
Mouse Models of Triplet Repeat Diseasesp. 3
Analysis of Triplet Repeat DNAs and RNAs
Analysis of Triplet Repeat Replication by Two-Dimensional Gel Electrophoresisp. 19
Genetic Analysis for Triplet Repeat Instability in Yeastp. 29
Detection and Isolation of Trinucleotide Repeat Expansions Using the RED Methodp. 47
Analysis of Unstable Triplet Repeats Using Small-Pool Polymerase Chain Reactionp. 61
Real-Time RT-PCR for CTG Repeat-Containing Genesp. 77
Detection and Analysis of Polyglutamine-Containing Proteins and Their Aggregates
Antibodies Against Huntingtin: Production and Screening of Monoclonals and Single-Chain Recombinant Formsp. 87
Using Antibodies to Analyze Polyglutamine Stretchesp. 103
Solubilization of Aggregates Formed by Expanded Polyglutamine Tract Expression in Cultured Cellsp. 129
Establishment of Animal and Cultured Cell Models for Trinucleotide Repeat Diseases
Caenorhabditis elegans as a Model System for Triplet Repeat Diseasesp. 141
Monitoring Aggregate Formation in Organotypic Slice Cultures From Transgenic Micep. 161
The CGG Repeat and the FMR1 Genep. 173
Analysis of CTG Repeats Using DM1 Model Micep. 185
Lentiviral-Mediated Gene Transfer to Model Triplet Repeat Disordersp. 199
Mouse Tissue Culture Models of Unstable Triplet Repeatsp. 215
In Vivo Analysis of Trinucleotide Repeat Diseases
Neurotransmitter Receptor Analysis in Transgenic Mouse Modelsp. 231
Chromatin Immunoprecipitation Technique for Study of Transcriptional Dysregulation in Intact Mouse Brainp. 261
Techniques for Thick-Section Golgi Impregnation of Formalin-Fixed Brain Tissuep. 277
Assessment of Impaired Proteasomal Function in a Cellular Model of Polyglutamine Diseasesp. 287
Assessment of In Vitro and In Vivo Mitochondrial Function in Friedreich's Ataxia and Huntington's Diseasep. 293
Triplet Repeats and DNA Repair: Germ Cell and Somatic Cell Instability in Transgenic Micep. 309
Oxidative Damage in Huntington's Diseasep. 321
Indexp. 335
Table of Contents provided by Rittenhouse. All Rights Reserved.

ISBN: 9781588292438
ISBN-10: 1588292436
Series: Methods in Molecular Biology
Audience: Professional
Format: Hardcover
Language: English
Number Of Pages: 342
Published: 18th June 2004
Publisher: Humana Press Inc.
Country of Publication: US
Dimensions (cm): 23.5 x 15.5  x 2.54
Weight (kg): 0.69