This chapter focuses on principles of isolation that seem likely to apply even when more demanding structural and functional tests of the product become available. It discusses the principles and then presents some examples of detailed procedures. Chromatin is usually isolated as fragments of chromosomal material, except for small plasmids and viral genomes, which may be isolated as intact minichromosomes. The main considerations in the isolation procedure are the nature of the starting material, the solution conditions, the methods of fragmentation and purification, and the criteria for preservation of the native state. The chief concern in the isolation of nuclei is with the loss of chromatin components. There is ample precedent for the leakage of nuclear proteins—for example, nucleoplasmin, which is localized to the nucleus in vivo, according to indirect immunofluorescence, but which is readily lost upon isolation. The importance of such components and the conditions needed for their retention during the isolation of nuclei will only be apparent when new structural and functional assays are available. Series: Methods in Enzymology (Book 170) Hardcover: 683 pages Publisher: Academic Press; 1 edition Language: English ISBN-10: 0121820718 ISBN-13: 978-0121820718 Product Dimensions: 1.5 x 6.5 x 9.8 inches Link Download http://nitroflare.com/view/11B140F675F22FFhttps://drive.google.com/drive/folders/1yLBzZ1rSQoNjmWeJTZ3WGQHg04L1
