Chromosome Microdissection and Cloning : A Practical Guide
This guide presents microdissection methodology, describing technical details, including "in-house secrets". The manual has been designed to aid the novice, as well as the experienced researcher, in setting up laboratories from "scratch" for the study of genetic abnormalities and chromosome aberrations. The role of those techniques in a number of new areeas, particularly in the human genome project, in gene identification in inheritable disorders, and in gene isolation in cancer research, is discussed.
- Spiral bound | 134 pages
- 196.85 x 234.95 x 9.91mm | 403.7g
- 01 Oct 1993
- Elsevier Science Publishing Co Inc
- Academic Press Inc
- San Diego, United States
- glossary, references, index
Table of contents
Part 1 Introduction to chromosome microdissection: chromosome organization; cloning DNA from chromosome fragments. Part 2 Preparation of chromosome for microdissection: critical aspects of chromosome preparation; reagents; equipment; protocols. Part 3 Methods of chromosome microdissection: methods; summary of chromosome microdissection and collection of DNA cloning. Part 4 Molecular cloning of microdissected chromosomal DNA: cloning DNA from microdissected chromosomal DNA fragments; direct cloning from microdissected chromosomal fragments; ligation of microdissected chromosomal DNA with plasmid vector or linker adapter and PCR amplification; PCR amplification of microdissected chromosomal DNA fragments by probing a complete recombinant library; analysis of recombinant clones derived from microdissected chromosomal DNA; determination of DNA insert size range; determination of the percentage of recombinant clones containing repeat and unique sequences; calculation of the percentage of microdissected DNA cloned; chromosomal localization of recombinant clones using fluorescent "in situ" hybridization. Part 5 Applications of chromosome microdissection: analysis of PCR product of microdissected chromosome fragments; recombinant DNA libraries from microdissected chromosome fragments; gene transfer using chromosome fragments.