Biotechnology: A Laboratory Course
- ISBN-10
- 0120845628
- ISBN-13
- 9780120845620
- Series
- Biotechnology
- Category
- Medical
- Pages
- 261
- Language
- English
- Published
- 1996
- Authors
- Guy A. Caldwell, Jeffrey M. Becker, Eve Ann Zachgo
Description
Asseptic technique and establishing pure cultures: the streak plate and culture transfer. Preparation of culture media. The growth curve. Isolation of plasmid DNA from escherichia coli: the mini-prep. Purification, concentration, and quantitation of DNA. Large-scale isolation of plasmid DNA by column chromatography. Amplification of a lacZ Gene fragment by the polymerase chain reaction. Restriction digestion and agarose gel electrophoresis. Southern transfer. Preparation, purification, and hybridization of probe. Transformation of saccharomyces cerevissiae. Isolation of plasmid from yeast and escherichia coli transformation. Protein assays. Qualitative assay for B-galactosidase in yeast colonies. Determination of B-galactosidase in permeabilized yeast cells. Assay of B-galactosidase in cell extracts. B-galactosidase purification. Western blot: probe of protein blot with antibody to B-galactosidase. Alternative protocols and experiments. Buffer solutions. Preparation of buffers and solutions. Properties of some common concentrated acids and bases. Use of micropipettors. List of cultures. Storage of cultures and DNA. Sterilization methods. Preparation of stock solutions for culture media. Growth in liquid medium. Determination of viable cells. Determination of cell mass. Determination of cell number. Nomenclature of strains. Glassware and plasticware. Preparation of tris and EDTA. Basic rules for handling enzymes. Effects of common contaminants on protein assays. Manufacturers' and distributors' addresses ...
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