Genomic Imprinting: Methods and Protocols

Description

Imprinted genes, many of which generally control growth and development, frequently lose their imprints during cancer progression, a loss that then plays a substantial role in uncontrolled tumor growth. Imprint instability also appears to be a major limitation to the success of mammalian cloning experiments. In Genomic Imprinting: Methods and Protocols, 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. The majority of protocols describe molecular techniques that allow examination of gene structure or expression in an allele-specific manner. 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. Incorporating a wealth of knowledge from leading exponents in the field, Genomic Imprinting: Methods and Protocols brings together all the essential molecular, genetic, and embryological methods commonly used in today's laboratories for the identification and analysis of imprinted genes.