On-Line Laboratory Manual 7th ed.


Lab Manual In Single PDF File (206 pages)

Table of Contents

About Genetic Engineering

Laboratory Resources

Digestion of DNA with Restriction Endonucleases

Agarose Gel Electrophoresis

Restriction Mapping

Experiment 1: Isolation of Escherichia coli Chromosomal DNA

Experiment 2: Determination of DNA Concentration and Purity by Ultraviolet Spectrophotometry

Experiment 3: The Chemical Nature of DNA

Experiment 4: DNA Sequence Specificity

Experiment 5: Activity of DNA Nucleases

Experiment 6: Restriction and Modificaion

Experiment 7: Analysis of Plasmid DNA by Restriction Digestion and Agarose Gel Electrophoresis

Experiment 8: Effect of Agarose Concentration on Migration of DNA Fragments

The First Lab Report

Experiment 9: Insertion of a Gene for Antibiotic Resistance from Bacillus subtilis into an Escherichia coli Plasmid

Experiment 10: Transformation of Escherichia coli With a Chimeric Plasmid

Experiment 11: Screening of Transformants for Chimeric Plasmids

Experiment 12: Large -Scale Purification of plasmids pRIT4501 and pRIT4502 by Cesium Chloride Density Gradient Centrifugation

Experiment 13: Large -Scale Purification of plasmids pRIT4501 and pRIT4502 by Qiagen Ion Exchange Column Chromatography

Experiment 14: Verification of Purified Plasmids

Experiment 15: Construction of a Restriction Map of Plasmids pRIT4501 and pRIT4502

The Second Lab Report

Experiment 16: Transformation of Plasmids pRIT4501 and pRIT4502 into Bacillus Subtilis

Experiment 17: Heterologous Gene Expression in Escherichia coli and Bacillus subtilis

The Third Lab Report

Experiment 18: Southern Blot Analysis of Plasmids pRIT4501 and pRIT4502

The Fourth Lab Report

Appendix I: Index of Reagent Formulas

Appendix II: DNA Structure

Appendix III - VII

Appendix III: Conversion Factors
Appendix IV: Buffer Dilutions
Appendix V: Bacillus subtilis Plasmid Prep by Qiagen Maxi Kit
Appendix VI: Effect of BAP Treatment on Transformation Frequencies
Appendix VII: Alpha Complementation

Appendix VIII: Density Gradient Centrifugation of DNA

Appendix IX: Bacteriophage Lambda

Appendix X: Technical Information

Appendix XI: Mrs. Rothman's Superstar Bars

Appendix XII: Cloning Methodology