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Genomic Imprinting: Methods and Protocols

Overview

Andrew Ward and a team of experienced researchers have brought together a collection of optimized classic and vanguard techniques for the identification and analysis of imprinted genes. Prools are included for identifying and cloning imprinted genes, for analyzing imprinted gene expression, for the study of DNA methylation and methylation-sensitive DNA-binding proteins, and for examining chromatin structure. There are also methods for the manipulation of mouse embryos to produce monoparental embryos and embryonic...

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Overview

Andrew Ward and a team of experienced researchers have brought together a collection of optimized classic and vanguard techniques for the identification and analysis of imprinted genes. Prools are included for identifying and cloning imprinted genes, for analyzing imprinted gene expression, for the study of DNA methylation and methylation-sensitive DNA-binding proteins, and for examining chromatin structure. There are also methods for the manipulation of mouse embryos to produce monoparental embryos and embryonic stem cells, and for the generation of transgenic mice with BAC, PAC,and YAC constructs. Each technique is described in step-by-step detail to ensure successful results.

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Editorial Reviews

From the Publisher
"...a multi-authored volume in which the methods of gene imprinting are discussed in detail in 25 chapters. The prools are very detailed with multiple schemes and figures." - Journal of Pediatric Endocrinology and Metabolism
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Product Details

  • ISBN-13: 9781617371646
  • Publisher: Springer-Verlag New York, LLC
  • Publication date: 11/10/2010
  • Series: Methods in Molecular Biology Series , #181
  • Edition description: Softcover reprint of hardcover 1st ed. 2002
  • Edition number: 1
  • Pages: 377
  • Product dimensions: 5.98 (w) x 9.02 (h) x 0.81 (d)

Table of Contents

Generation of Monoparental Embryos for Investigation into Genomic Imprinting
Wendy L. Dean, Gavin Kelsey, and Wolf Reik

Deriving and Propagating Mouse Embryonic Stem Cell Lines for Studying Genomic Imprinting
Jeffrey R. Mann

Balanced Translocations for the Analysis of Imprinted Regions of the Mouse Genome
Anne C. Ferguson-Smith, Maxine Tevendale, Pantelis Georgiades, and Valerie Grandjean

Production of YAC Transgenic Mice by Pronuclear Injection
Justin F.-X. Ainscough, Rosalind M. John, and Sheila C. Barton

A Transgenic Approach to Studying Imprinted Genes: Modified BACs and PACs
Rosalind M. John, Justin F.-X. Ainscough, and Sheila C. Barton

Methylation-Sensitive Genome Scanning
Izuho Hatada and Tsunehiro Mukai

Subtraction-Hybridization Method for the Identification of Imprinted Genes
Fumitoshi Ishino, Yoshimi Kuroiwa, Naoki Miyoshi, Shin Kobayashi, Takashi Kohda, and Tomoko Kaneko-Ishino

Identification of Imprinted Loci by Methylation: Use of Methylation- Sensitive Representational Difference Analysis (Me-RDA)
Rachel J. Smith and Gavin Kelsey

Ribonuclease Protection
Joanne L. Thorvaldsen and Marisa S. Bartolomei

Quantitative RT-PCR-Based Analysis of Allele-Specific Gene Expression
Judith Singer-Sam and Chunguang Gao

Allele-Specific In Situ Hybridization (ASISH)
Rolf Ohlsson, Kristian Svensson, Hengmi Cui, Helena Malmikumpu, and Gail Adam

RNA-FISH to Analyze Allele-Specific Expression
Giovanna Braidotti

Flow Cytometry and FISH to Investigate Allele-Specific Replication Timing and Homologous Association of Imprinted Chromosomes
Janine LaSalle and Marc Lalande

Southern Analysis Using Methyl-Sensitive Restriction Enzymes
Tom Moore

A PCR-Based Method for Studying DNA Methylation
Mira Ariel

Bisulfite-Based Methylation Analysis of Imprinted Genes
Sabine Engemann, Osman El-Maarri, Petra Hajkova, Joachim Oswald, and Joern Walter

Direct Analysis of Chromosome Methylation
Déborah Bourc'his and Evani Viegas-Péquignot

In Vitro Methylation of Predetermined Regions in Recombinant DNA Constructs
Ilse Van den Wyngaert, Roger L. P. Adams, and Stefan U. Kass

In Vitro Methylation of Specific Regions in Recombinant DNA Constructs by Excision and Religation
Ghislaine Dell, Marika Charalambous, and Andrew Ward

Detection of Methyl-Sensitive DNA-Binding Proteins with Possible Involvement in the Imprinting Phenomenon
Kerstin Otte and Björn Rozell

Probing Chromatin Structure with Nuclease Sensitivity Assays
Richard I. Gregory, Sanjeev Khosla, and Robert Feil

Examining Histone Acetylation at Specific Genomic Regions
Ji-Fan Hu and Andrew R. Hoffman

Purification of the MeCP2/Histone Deacetylase Complex from Xenopus laevis
Peter L. Jones, Paul A. Wade, and Alan P. Wolffe

Reconstitution of Chromatin In Vitro
Kiyoe Ura and Yasufumi Kaneda

Genomic Imprinting in Plants
Rinke Vinkenoog, Melissa Spielman, Sally Adams, Hugh G. Dickinson, and Rod J. Scott

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