Genomic imprinting : methods and protocols / edited by Andrew Ward.

Format:

Book

Contributor:

Ward, Andrew, 1964-

Series:

Methods in molecular biology ; v. 181.

Methods in molecular biology ; v. 181

Language:

English

Subjects (All):

Genomic imprinting--Laboratory manuals.

Genomic imprinting.

Genomic Imprinting.

Medical Subjects:

Genomic Imprinting.

Genre:

Laboratory manuals.

Physical Description:

xii, 377 pages : illustrations ; 24 cm.

Place of Publication:

Totowa, NJ : Humana Press, 2002.

Summary:

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. Protocols 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.

Contents:

1 Generation of Monoparental Embryos for Investigation into Genomic Imprinting / Wendy L. Dean, Gavin Kelsey, Wolf Reik 1

2 Deriving and Propagating Mouse Embryonic Stem Cell Lines for Studying Genomic Imprinting / Jeffrey R. Mann 21

3 Balanced Translocations for the Analysis of Imprinted Regions of the Mouse Genome / Anne C. Ferguson-Smith, Maxine Tevendale, Pantelis Georgiades, Valerie Grandjean 41

4 Production of YAC Transgenic Mice by Pronuclear Injection / Justin F.-X. Ainscough, Rosalind M. John, Sheila C. Barton 55

5 A Transgenic Approach to Studying Imprinted Genes: Modified BACs and PACs / Rosalind M. John, Justin F.-X. Ainscough, Sheila C. Barton 67

6 Methylation-Sensitive Genome Scanning / Izuho Hatada, Tsunehiro Mukai 83

7 Subtraction-Hybridization Method for the Identification of Imprinted Genes / Fumitoshi Ishino, Yoshimi Kuroiwa, Naoki Miyoshi, Shin Kobayashi, Takashi Kohda, Tomoko Kaneko-Ishino 101

8 Identification of Imprinted Loci by Methylation: Use of Methylation-Sensitive Representational Difference Analysis (Me-RDA) / Rachel J. Smith, Gavin Kelsey 113

9 Ribonuclease Protection / Joanne L. Thorvaldsen, Marisa S. Bartolomei 133

10 Quantitative RT-PCR-Based Analysis of Allele-Specific Gene Expression / Judith Singer-Sam, Chunguang Gao 145

11 Allele-Specific In Situ Hybridization (ASISH) / Rolf Ohlsson, Kristian Svensson, Hengmi Cui, Helena Malmikumpu, Gail Adam 153

12 RNA-FISH to Analyze Allele-Specific Expression / Giovanna Braidotti 169

13 Flow Cytometry and FISH to Investigate Allele-Specific Replication Timing and Homologous Association of Imprinted Chromosomes / Janine LaSalle, Marc Lalande 181

14 Southern Analysis Using Methyl-Sensitive Restriction Enzymes / Tom Moore 193

15 A PCR-Based Method for Studying DNA Methylation / Mira Ariel 205

16 Bisulfite-Based Methylation Analysis of Imprinted Genes / Sabine Engemann, Osman El-Maarri, Petra Hajkova, Joachim Oswald, Joern Walter 217

17 Direct Analysis of Chromosome Methylation / Deborah Bourc'his, Evani Viegas-Pequignot 229

18 In Vitro Methylation of Predetermined Regions in Recombinant DNA Constructs / Ilse Van den Wyngaert, Roger L. P. Adams, Stefan U. Kass 243

19 In Vitro Methylation of Specific Regions in Recombinant DNA Constructs by Excision and Religation / Ghislaine Dell, Marika Charalambous, Andrew Ward 251

20 Detection of Methyl-Sensitive DNA-Binding Proteins with Possible Involvement in the Imprinting Phenomenon / Kerstin Otte, Bjorn Rozell 259

21 Probing Chromatin Structure with Nuclease Sensitivity Assays / Richard I. Gregory, Sanjeev Khosla, Robert Feil 269

22 Examining Histone Acetylation at Specific Genomic Regions / Ji-Fan Hu, Andrew R. Hoffman 285

23 Purification of the McCP2/Histone Deacetylase Complex from Xenopus Iaevis / Peter L. Jones, Paul A. Wade, Alan P. Wolffe 297

24 Reconstitution of Chromatin In Vitro / Kiyoe Ura, Yasufumi Kaneda 309

25 Genomic Imprinting in Plants / Rinke Vinkenoog, Melissa Spielman, Sally Adams, Hugh G. Dickinson, Rod J. Scott 327.

Notes:

Includes bibliographical references and index.

ISBN:

089603741X

OCLC:

46641762