YAC Libraries: A User's Guide

Overview

An indispensable guide to an emerging technology, this unique book introduces detailed methods for the construction, screening, and use of DNA sequences cloned into yeast artificial chromosome (YAC) vectors and the applications of YACs to gene hunting. Also covered are the reconstruction of large genes and the analysis of YACs by insertion back into mammalian cells—a method that may lead to the therapeutic use of YAC clones. The methods described are drawn from work on human DNA, but are universal to yeast ...

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Overview

An indispensable guide to an emerging technology, this unique book introduces detailed methods for the construction, screening, and use of DNA sequences cloned into yeast artificial chromosome (YAC) vectors and the applications of YACs to gene hunting. Also covered are the reconstruction of large genes and the analysis of YACs by insertion back into mammalian cells—a method that may lead to the therapeutic use of YAC clones. The methods described are drawn from work on human DNA, but are universal to yeast artificial chromosomes used in other organisms. YAC Libraries is the third volume in the UWBC Biotechnical Resource Series with Dr. Richard Burgess, Director of the University of Wisconsin Biotechnology center, as Series Editor. The series is dedicated to providing practical guidance to researchers exploring new technologies and services.

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Editorial Reviews

Booknews
For researchers who have not used it before, introduces the methods for constructing, screening, and using DNA sequences cloned into yeast artificial chromosome vectors for hunting genes. Among the topics are vectors and host strains, localizing genes on metaphase chromosomes, and cloning human disease genes. Annotation c. Book News, Inc., Portland, OR (booknews.com)
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Product Details

Meet the Author

Baylor College of Medicine

Washington University School of Medicine

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Table of Contents

Foreword
Introduction
1 Vectors and Host Strains for Cloning and Modification of Yeast Artificial Chromosomes 1
2 Yeast Spheroplast Transformation with Large DNA 33
3 Large Insert Yeast Artificial Chromosome Libraries 43
4 Isolation of YAC Clones from Genomic Libraries by Southern Blot Hybridization 57
5 Generation of YAC Contigs by Walking 93
6 The Use of YACs to Identify Expressed Sequences: cDNA Screening Using Total YAC Insert 113
7 YAC Clones as an Integral Component in the Positional Cloning of Human Disease Genes: Isolation of the APC Gene 127
8 Transfer of YAC DNA into Mammalian Cells and Transgenic Mice 143
9 Fluorescent in Situ Hybridization to Metaphase Chromosomes with YACs 165
10 YAC Breeding to Reconstitute Large Loci 181
Appendix: Growth Media 203
Index 207
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