Gene Isolation and Mapping Protocols / Edition 1

by Jacqueline Boultwood

Overview

Over the past 10 years great progress has been made in the development of efficient techniques for both gene isolation and mapping. The identifica- tion and isolation of transcribed sequences from large chromosomal regions are central to the human genome mapping project. Techniques for isolating novel cDNAs have applications both in the overall construction and integra- tion of long-range physical and transcription maps and in the identification of disease genes. A number of different techniques for the isolation of cDNAs from mam- malian genomes have been developed, including screening "zoo" blots, the use of large genomic clones (YACs or cosmids) for hybridization against cDNA libraries, and CpG island mapping. More recently two highly efficient tech- niques have been introduced: exon trapping, based on the presence of exon splice sites, and direct selection, based on the enrichment of selected cDNAs using immobilized YACs or cosmids. Leading researchers in the field have contributed chapters detailing the practical procedures for these and other widely used methods. The most rapid progress presently being made in the field of gene isolation concerns the partial sequencing of cDNA clones from one or both ends to produce expressed sequence tags (ESTs). Indeed, by Octo- ber 1995, the EST division of Genbank (dbEST) contained a total of approxi- mately 270,000 human EST sequences accounting for almost half the number of sequence entries in Genbank.

Product Details

ISBN-13:	9780896033825
Publisher:	Springer-Verlag New York, LLC
Publication date:	10/19/1996
Series:	Methods in Molecular Biology , #68
Edition description:	1997
Pages:	318
Product dimensions:	5.98(w) x 9.02(h) x 0.03(d)

About the Author

Boultwood, Jacqueline (John Radcliffe Hospital, Oxford)

The contributors represent the specialties of molecular biology, biochemistry, hematology/oncology, genetics, microbiology, pediatrics, cell biology, bioinformatics, immunochemistry, and psychiatry. Most are from universities, academic hospitals, and research institutes in the U.K., U.S., France, and the Netherlands. Institutions prominently represented include Oxford, Leiden Univ, Univ of Texas Southwestern Med Ctr, Univ of Chicago, MD Anderson Cancer Ctr, Johns Hopkins, NIH, and Vanderbilt.

Table of Contents

Gene Mapping Goes from FISH to Surfing the Net.- Linkage Analysis of Genetic Disorders.- Gene Ordering and Localization by Linkage Analysis.- Gene Mapping Using Somatic Cell Hybrids.- Gene Mapping by FISH.- Probe Ordering and Distancing by FISH.- Physical Mapping by Pulsed-Field Gel Electrophoresis.- Construction and Use of YAC Contigs in Disease Regions.- Construction and Use of Cosmid Contigs.- Use of Dinucleotide Polymorphism Analyses in Physical Mapping.- Mapping Expressed Sequence Tags (ESTs) by Multiplexing PCR Reactions from Hybrid Cell Panels and Detecting Fluorescently Labeled Products.- Gene Isolation by Exon Trapping.- Isolation of Coding Sequences from Genomic Regions Using Direct Selection.- Isolation of cDNAs Using the YAC Hybridization Screen Method.- Detection and Isolation of Differentially Expressed Genes by Differential Display.- Chemical Crosslinking Subtraction (CCLS).- Gene Mapping and Isolation.- Software for Genetic Linkage Analysis.- Exon Detection by Similarity Searches.

Editorial Reviews

Reviewer: Roger W. Melvold, PhD (Northwestern University Feinberg School of Medicine)
Description: This new addition to the Methods in Molecular Biology series describes the techniques useful in the isolation and mapping of genes at the molecular levels.
Purpose: The volume opens with a brief overview of progress in the field during the recent years. The chapters are logically arranged; for example, the chapter on "Gene Ordering and Localization by Linkage Analysis" is preceded by a chapter describing the genetic and mathematical basis for linkage analysis.
Audience: The volume is directed at geneticists and molecular biologists involved in the area, whether in research laboratories or diagnostic laboratories. Thirty-five contributors have participated in the 19 chapters.
Features: Areas covered include the use of somatic cell hybrids, YAC chromosome technology, and fluoresence in situ hybridization. Also provided are chapters covering a variety of physical mapping techniques at the nucleotide level. Several of the chapters contain excellent and appropriate black-and-white illustrations or photomicrographs. The technique chapters are organized along a consistent format (Introduction, Materials, Methods, Notes, and References). Of particular interest are multiple chapters describing the availability and application of electronic databases for identifying matching sequences and for linkage analysis.
Assessment: The volume is a useful addition to the libraries of individuals and institutions engaged in relevant laboratory activities.

Doody's Review Service

...a useful addition to the libraries of individuals and institutions... Three Stars. - Doody's Health Sciences Book Review Journal

"...this series of excellent and detailed lab manuals....of general use to all those mapping and isolation genes."-SGM Quarterly

From the Publisher

3 Stars from Doody

This new addition to the Methods in Molecular Biology series describes the techniques useful in the isolation and mapping of genes at the molecular levels. The volume opens with a brief overview of progress in the field during the recent years. The chapters are logically arranged; for example, the chapter on "Gene Ordering and Localization by Linkage Analysis" is preceded by a chapter describing the genetic and mathematical basis for linkage analysis. The volume is directed at geneticists and molecular biologists involved in the area, whether in research laboratories or diagnostic laboratories. Thirty-five contributors have participated in the 19 chapters. Areas covered include the use of somatic cell hybrids, YAC chromosome technology, and fluoresence in situ hybridization. Also provided are chapters covering a variety of physical mapping techniques at the nucleotide level. Several of the chapters contain excellent and appropriate black-and-white illustrations or photomicrographs. The technique chapters are organized along a consistent format (Introduction, Materials, Methods, Notes, and References). Of particular interest are multiple chapters describing the availability and application of electronic databases for identifying matching sequences and for linkage analysis. The volume is a useful addition to the libraries of individuals and institutions engaged in relevant laboratory activities.

Roger W. Melvold