Practical Methods in Molecular Biology
Springer-Verlag New York Inc.
978-1-4612-5958-9 (ISBN)
This volume has evolved from a laboratory methods book that one of us first compiled nearly fifteen years ago. Since that time the book has undergone many minor revisions in order to include new methods and updated versions of older methods. The result has been an increasingly useful and more widely circulated book. However, the recent series of technological explosions generally lumped together under the name of "recombinant DNA technology" has been a turning point in the evolution of this previously underground publication. Minor revisions will no longer do. To keep the book useful we have had to make major revi sions and additions. The result is a dramatically expanded book that should be more useful to more people. The larger size and wider usefulness of the book have made this more formal publication seem a reasonable step to take. One of the reasons that this volume should be useful to many people is that it includes only procedures that have been used repeatedly by us and that have proven highly reliable both to ourselves and to others in our laboratories.
1 Using E. coli.- Strains.- Cell Growth.- Measuring Cell Density.- Growing Large Quantities of Cells.- Opening Cells.- Radiolabelling Cells.- Nitrosoguanidine Mutagenesis.- Penicillin Selection.- Curing Cells of F-Factors.- Making Cells Streptomycin-Resistant.- Crossing recA into Cells.- Phage P1 Transduction of Genetic Markers.- Large-Scale Genetic Crosses.- Using Transposons in Strain Construction.- 2 Bacteriophage Lambda.- Two Useful Mutants.- Titering.- Growing Plate Stocks.- Large-Scale Growth in Liquid.- Purification.- Genetic Crosses.- Scoring Plaques.- Making Strains Lambda-Resistant.- Testing Colonies for the Ability to Grow Lambda.- Making Lysogens.- Testing Lysogen Candidates.- Streaking for Single Plaques.- Selecting Deletions.- 3 Enzyme Assays.- ?-Galactosidase.- RNA Polymerase.- Arabinose Isomerase.- Lysozyme.- Ribulokinase.- E. coli-Coupled Transcription-Translation System.- 4 Working with Proteins.- Ammonium Sulfate Precipitation of Proteins.- Removing Nucleic Acids by Phase Partition.- Columns, Fraction Collectors, and Plumbing.- Ion Exchange Chromatography and Gel Filtration.- Determining Protein Concentration.- Concentrating Protein Solutions.- Stabilizing Proteins.- Polyacrylamide Gel Electrophoresis of Proteins.- 5 Working with Nucleic Acids.- Measuring Nucleic Acid Concentration and Purity.- Storing DNA.- Cleaning DNA.- Precipitating DNA with Ethanol.- TCA Precipitation Assay.- Precipitation and Size Fractionation of DNA with Poly-ethylene Glycol.- Isolating E. coli DNA.- Isolating Lambda DNA.- Isolating Plasmid DNA.- Large-Scale Plasmid Isolation.- Isolating Drosophila DNA.- Preparing Nucleoside Triphosphate Solutions.- Chromatographic Analysis of Nucleosides.- Gel Electrophoresis of DNA.- Mapping Restriction Endonuclease Sites on DNA.- 6Constructing and Analyzing Recombinant DNA.- Joining the Ends of DNA Molecules.- E. coli Transformation with Plasmid DNA.- Storing Strains that Contain Plasmids.- Cycloserine Selection of Recombinant Plasmids.- In Vitro Radiolabelling of DNA and RNA.- General Aspects of Nucleic Acid Hybridization Reactions.- Screening Recombinant DNA Clones by Nucleic Acid Hybridization.- Isolating DNA from a Single Colony.- Southern Transfers.- Selecting RNA Complementary to a DNA.- In Vitro Translation Systems from Higher Organisms.- Purifying Total and Polysomal RNA.- Purification of Poly-A+ (mRNA-Enriched) RNA.- RNA Size Fractionation by Sucrose Gradient Centrifugation.- 7 Assorted Laboratory Techniques.- Glass and Plastic Containers.- Siliconizing Glassware.- Washing Pipettes.- pH Meters.- Buffers (Tris, Phosphate, Good Buffers, Cacodylate).- Beckman Ultracentrifuges.- Drawing Figures.- Slides and Negatives.- Film Sensitometry.- Autoradiography and Fluorography.- Dialysis Tubing.- Distilling Phenol.- Recovering Used CsCl.- Sources of Chemicals.- Hazards and Cautions.- Appendix I Commonly Used Recipes.- Appendix II Useful Numbers.
| Zusatzinfo | XIV, 220 p. |
|---|---|
| Verlagsort | New York, NY |
| Sprache | englisch |
| Maße | 210 x 280 mm |
| Themenwelt | Sachbuch/Ratgeber ► Natur / Technik ► Garten |
| Naturwissenschaften ► Biologie ► Genetik / Molekularbiologie | |
| Naturwissenschaften ► Biologie ► Mikrobiologie / Immunologie | |
| Naturwissenschaften ► Biologie ► Zellbiologie | |
| Schlagworte | Biology • Molekularbiologie |
| ISBN-10 | 1-4612-5958-4 / 1461259584 |
| ISBN-13 | 978-1-4612-5958-9 / 9781461259589 |
| Zustand | Neuware |
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