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Recombinant DNA laboratory manual /
The latest edition of this introductory benchtop manual is up-to-date, affordable, and easy-to-follow. This text is perfect for your two-quarter or one semester course in Recombinant DNA Techniques and is specifically designed to lead your student or technician, who is a newcomer to molecular biolog...
| Clasificación: | Libro Electrónico |
|---|---|
| Autor principal: | |
| Otros Autores: | |
| Formato: | Electrónico eBook |
| Idioma: | Inglés |
| Publicado: |
San Diego :
Academic Press,
�1992.
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| Edición: | Rev. ed. |
| Temas: | |
| Acceso en línea: | Texto completo |
Tabla de Contenidos:
- Front Cover; Copyright Page; Recombinant DNA Laboratory Manual; Dedication; Table of Contents; PREFACE; SCHEDULE OF LABORATORY EXERCISES; Lab I. BACTERIAL GROWTH PARAMETERS; Day 1. Measuring Bacterial Cell Growth; Day 2. Plotting Cell Growth Data; Addendum; Lab II. Isolation and Analysis of Bacterial and Drosophila Chromosomal DNA; Day 1. Isolation and Purification of E. coli Chromosomal DNA; Day 2. Determination of the Concentration and Purity of DNA by UV Spectroscopy; Day 3. Restriction Endonuclease Digestion of Chromosomal DNA
- Day 4. Agarose Gel of Chromosomal DNA Restriction Endonuclease DigestionsDay 5. Staining and Photography of Agarose Gel of Chromosomal DNA Restriction Endonuclease Digestions; Lab III. Plasmid DNA Isolation and Agarose Gel Analysis; Day 1. Isolation of Plasmid DNA by the Alkaline-Detergent Method- A Miniprep Procedure; Day 2. Agarose Gel Electrophoresis of Undigested Plasmid DNA; Lab IV. Introduction of DNA into Cells; Day 1. Production of Frozen Competent Cells; Day 2. Transformation of LE392 with pBR329 DNA Isolated from HB101 ::Tn5; Lab V. Tn5 Mutagenesis of PBR329
- Day 1. Marker Screening: Divide Transformants into Tcs and Tcr ClassesDay 2. Purification of Tcs and Tcr Clones; Day 3. Isolation of Plasmid DNA by the Alkaline-Detergent Method and Determination of Recovery by Agarose Gel Electrophoresis; Day 4. Restriction Mapping of the Tn5 Inserts using Pstl and EcoRl; Day 5. Agarose Gel of Plasmid DNA Restriction Endonuclease Digestions; Lab VI. DNA Cloning in M13; Day 1. Isolation of Restriction Fragment from an Agarose Gel; Day 2. Estimation of Recovery of Restriction Fragment and Isolation of M13mp 19 RF DNA
- Day 3. EcoRl Digestion of M13mpl9 RF DNA and Treatment with Alkaline PhosphataseDay 4. Removing the Phosphatase and EcoRl and Analysis of the EcoRl Digest of M13mpl9 RF DNA; Day 5. Ligation; Ligation of EcoRl Digested M13mpl9 RF DNA and the Purified pRSG192 EcoRl Fragment; Day 6. Transfection of XL 1-Blue with Ligation Mixtures; Day 7. Plaque Purification; Day 8. Isolation of Colorless Plaques That Contain the chb Gene and Growth of Recombinant Bacteriophage; Lab VII. DNA Sequencing; Day 1. Isolation of Recombinant M13mpl9 RF and ss DNA
- Day 2. Restriction Digestion and Gel Electrophoresis of Recombinant Phage to Determine Orientation of InsertEstimating the Recovery of ss DNA using Agarose Gel Electrophoresis; Day 3. Staining Gel of Pstl Restriction Fragments; Template Annealing and Dideoxy Sequencing; Day 4. Electrophoretic Separation of Sequencing Reactions; Day 5. Developing Autoradiogram and Reading DNA Sequence; Lab VIII. DNA Gel Blotting, Probe Preparation, Hybridization, and Hybrid Detection; Day 0. Agarose Gel Electrophoresis; Day 1. Gel Blotting; Day 2. Baking the Blot, Nick Translation, and Biotinylation of DNA