YAC libraries : a user's guide / edited by David L. Nelson, Bernard H. Brownstein.

New York : W.H. Freeman and Co., [1994], ©1994.

xx, 211 pages : illustrations ; 23 cm.

Copy 1 Donor: Wellcome Trust Sanger Institute.

Vectors and host strains for cloning and modification of yeast artificial chromosomes -- YAC cloning vectors -- Accessory vectors for modification of YACs -- Host strains for cloning and manipulation of YACs -- Yeast spheroplast transformation with large DNA -- Aqueous transformations -- Isolation of high molecular weight yeast DNA in agarose plugs -- Inverse PCR of the ends of YAC inserts -- Inverse PCR for identifying intron/exon boundaries -- Subcloning YAC fragments into a lambda vector -- Transfer of YAC DNA into mammalian cells and transgenic mice -- Mammalian selectable markers -- Fusion of yeast spheroplasts with mammalian cells -- Solutions -- Preparation of yeast spheroplasts -- Fusion protocol -- Transformation with total yeast DNA -- Solutions -- Preparation of high molecular weight yeast DNA by centrifugation through a sucrose gradient -- Lipofectin-mediated DNA transfer -- Transformation with gel-purified YAC DNA -- Preparation of YAC DNA by agarasing a gel slice -- Microinjection into tissue-culture cells -- Lipofection into mammalian cells.

(cont) Concentration with a filter unit (up to 600 kb remains intact) -- Microinjection of oocytes for generation of transgenic mice -- Fluorescent in situ hybridization to metaphase chromosomes with YACs -- Current fluorescent in situ hybridization (FISH) -- Implications of FISH for genome mapping -- FISH using YACs -- YAC breeding to reconstitute large loci -- Materials and methods -- Growth media.

Includes bibliographical references and index.