Unraveling DNA Molecular Biology for the Laboratory

ISBN-10: 0132700344

ISBN-13: 9780132700344

Edition: 1st 1997

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Description: This innovative mannual introduces students to all of the basic techniques of modern molecular biology using an integrated series of laboratory exercises that involve the cloning and analysis of the bioluminescene genes.

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Book details

List price: $123.80
Edition: 1st
Copyright year: 1997
Publisher: Benjamin-Cummings Publishing Company
Publication date: 1/22/1997
Binding: Paperback
Pages: 400
Size: 8.50" wide x 11.00" long x 0.75" tall
Weight: 2.156
Language: English

Introductory Techniques
Introduction to the Laboratory: Basic Equipment and Bacteriological Techniques
Preparation of Media and Reagents Used in Molecular Biology
Isolation of Luminescent Bacteria from Natural Sources
Restriction Digestion and Agarose Gel Electrophoresis of DNA
DNA Isolation and Analysis
Isolation of Chromosomal DNA from Vibrio fischeri
Large-Scale Purification of Plasmid DNA
Spectrophotometric Analysis of DNA
Cloning the Lux Operon
Restriction Digestion of Vibrio fischeri Genomic DNA and Plasmid Vector
Quantification of Genomic DNA by Fluorometry and Agarose Plate Fluorescence
Ligation of Restriction Fragments of Vibrio fischeri DNA to Plasmid Vector
Preparation of Competent Escherichia coli DH5
Transformation of Competent Escherichia coli DH5 with Recombinant Plasmids
Screening the Vibrio fischeri Genomic Library for Light Producing Clones
Restriction Mapping and Southern Blotting
Small-Scale Plasmid Isolations (Mini-preps) from Bioluminescent Clones
Restriction Mapping of Plasmids from Bioluminescent Clones
Southern Blotting and Hybridization to Detect the luxA Gene
Subcloning the Lux A Gene
Restriction Digestion of lux Plasmids and Cloning Vector for Subcloning luxA
Gel Purification of DNA Restriction Fragments Containing luxA
Subcloning luxA into A Plasmid Vector
Transformation of Competent Escherichia coli DH5 with Subcloned DNA
Colony Hybridization to Screen for luxA Subclones
Small-Scale Plasmid Isolations (Mini-Preps) from luxA Clones
Advanced Techniques
Amplification of luxA from Natural Isolates by the Polymerase Chain Reaction (PCR)
Southern Blotting and Hybridization of PCR Products
DNA Sequencing of lux Genes from Plasmid Templates
Computer Analysis of DNA Sequences Using the World Wide Web
Mapping the Vibrio fischeri Genome by Pulsed Field Gel Electrophoresis
Independent Projects in Molecular Biology
Appendices
The Metric System and Units of Measure
Centrifugation
Spectrophotometry
Agarose and Polyacrylamide Gel Electrophoresis
Methylene Blue Staining of Agarose Gels
Nucleic Acid Hybridization
Alcohol Precipitation of Nucleic Acids
Care and Handling of Enzymes
Restriction Endonucleases
Enzymes Used in Molecular Biology
Maps of Cloning Vectors and Bacteriophage Lambda
Proper Handling and Disposal of Hazardous Materials
Procedures and Precautions for the Use of Radioisotopes
Equipment and Supplies
Media and Reagents
Bacterial Strains
Lists of Suppliers
Recommended References
Restriction Mapping Problems
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