Detail Chromatin Immunoprecipitation and Microarray-Based Analysis of Protein Location (in Nature Protocols Vol.1 No.2) Bibliografi Author: Lee, Tong Ihn ; Johnstone, Sarah E. ; Young, Richard A. Topik: Chromatin Immunoprecipitation (ChIP); Microarray; ChIP-Chip Bahasa: (EN )    Edisi: Jul 2006     Penerbit: Nature Publishing Group     Tempat Terbit: New York    Tahun Terbit: 2006     Jenis: Article - diterbitkan di jurnal ilmiah internasional Fulltext: RAYoung.pdf (635.96KB; 0 download) [Informasi yang berkaitan dengan koleksi ini di internet] Abstract Genome-wide location analysis, also known as ChIP-Chip, combines chromatin immunoprecipitation and DNA microarray analysis to identify protein-DNA interactions that occur in living cells. Protein-DNA interactions are captured in vivo by chemical crosslinking. Cell lysis, DNA fragmentation and immunoaffinity purification of the desired protein will co-purify DNA fragments that are associated with that protein. The enriched DNA population is then labeled, combined with a differentially labeled reference sample and applied to DNA microarrays to detect enriched signals. Various computational and bioinformatic approaches are then applied to normalize the enriched and reference channels, to connect signals to the portions of the genome that are represented on the DNA microarrays, to provide confidence metrics and to generate maps of protein-genome occupancy. Here, we describe the experimental protocols that we use from crosslinking of cells to hybridization of labeled material, together with insights into the aspects of these protocols that influence the results. These protocols require approximately 1 week to complete once sufficient numbers of cells have been obtained, and have been used to produce robust, high-quality ChIP-chip results in many different cell and tissue types. Opini Anda Klik untuk menuliskan opini Anda tentang koleksi ini! Lihat Sejarah Pengadaan  Konversi Metadata   Kembali

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