Biotechnology: a laboratory course
Record keeping and safety rules; Measurement of pH; Use of micropipettors and spectrophotometers; Aseptic technique: transfering a culture; Establishing a pure culture: the streak plate; Preparation of culture media; The growth curve; Isolation of plasmid DNA from Escherichia coli: the mini-prep; Purification, concentration, and quantitation of DNA; Isolation of plasmid DNA: the maxi-prep; Restriction digestion and agarose gel electrophoresis; Southern transfer; Preparation, purification, and hybridization of probe; Transformation of Saccharomyces cerevisiae; Transformation of Escherichia coli by plasmid DNA; Protein assays; B-galactosidase assay; Determination of B-galactosidase in permeabilized yeast cells; Assay of B-galactosidase in cell extracts; B-galactosidase purification.
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Measurement of pH
Use of Micropipettors and Spectrophotometers
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20 minutes 3-galactosidase agar plates agarose ammonium sulfate ampicillin Appendix aseptic Autoclave bacteria Beckman biotinylated Bradford protein assay bromphenol blue buffer pH cell extract cells/ml centrifuge tube cerevisiae YNN281 chromatography cloning colonies column concentration containing dilution discard distilled water DNA fragments EDTA electrophoresis Escherichia coli EtBr ethanol Ethidium bromide Exercise filter flaming flask gel filtration gently glass growth hybridization Incubator at 30°C inoculating loop Instructor's Note Isolation of Plasmid laboratory LB broth lntroduction lysozyme maxi-prep medium method mg/ml microcentrifuge tube micropipettor mini-prep ml of distilled molecular weight molecules mutants NaCl NaOH nitrocellulose Pasteur pipet pellet petri plates phase phenol plasmid plasmid DNA prepare probe Procedure purification reaction reagent Reagents/Supplies remove restriction digests resuspend room temperature Saccharomyces cerevisiae Safety Note sample sodium Spectrophotometer step sterile stock solution streak plate supernatant test tube transfer transformation UV light volume Vortex water bath xg/ml yeast