Manipulation and expression of recombinant DNA : a laboratory manual /
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| Author / Creator: | Carson, Sue (Susan) |
|---|---|
| Edition: | 2nd ed. |
| Imprint: | Burlington, MA : Elsevier Academic Press, ©2006. |
| Description: | 1 online resource (xvi, 151 pages) : illustrations (some color) |
| Language: | English |
| Subject: | |
| Format: | E-Resource Book |
| URL for this record: | http://pi.lib.uchicago.edu/1001/cat/bib/11148737 |
Table of Contents:
- Cover; Contents; Preface; Acknowledgments; Note to Instructors; Instrumentation; Nomenclature; Introduction; Conceptual Outline for Experiments; Experimental Procedures; Laboratory Safety; General Operating Procedures; Emergency Contact Information; Part I; Manipulation of DNA; Lab Session 1; Getting Oriented; Practicing with Micropipettes; Station Checklist; Micropipetting; Laboratory Exercise; Preparing BSA Dilutions; Performing a Nitrocellulose Spot Test; Lab Session 2; Large-Scale Purification of Plasmid DNA; Introduction; Laboratory Exercises
- Alkaline Lysis and Anion Exchange ChromatograhyDNA Quantification; Lab Session 3; Preparation of Expression Vector DNA (pET-41a(+), a GST Fusion Protein Vector); Introduction; Laboratory Exercises; Restriction Digestion of Vector (pET41a) Restriction Enzyme Digestions; Agarose Gel Electrophoresis; Cleaning DNA Using a Spin Column; Lab Session 4; Preparation of Insert DNA (egfp); Introduction; Laboratory Exercises; Restriction Digestion of pEGFP-N1; Isolation of egfp DNA from Agarose; Lab Session 5; Preparation of Transformation-Competent Cells and Control Transformation; Introduction
- Laboratory Exercises Preparation of Chemically Competent Cells by Calcium Chloride Treatment; Transformation Control; Lab Session 6; DNA Ligation and Transformation of Escherichia coli; Introduction; Laboratory Exercises; Ligations and Ligation Controls; Divalent Cation-Mediated Transformation; Electrophoresis of Ligation Reactions; Part II; Screening Transformants; Lab Session 7; Colony Hybridizations; Lab Session 7A; Interim Laboratory Session; Introduction; Laboratory Exercises; Counting Transformants; Replica Plating; Lab Session 7B; Colony Hybridization: DNA Probe; Introduction
- Laboratory Exercises Colony Hybridization with an egfp DNA Probe: Part 1; Labeling of DNA Probe by PCR Using Digoxigenin-11-dUTP; Lab Session 7C; Colony Hybridization: Monoclonal Antibody Probe; Introduction; Laboratory Exercise; Colony Hybridization with an a-GFP Monoclonal Antibody Probe: Part 1; Lab Session 8; Completion of Colony Hybridization with DNA Probe; Introduction; Laboratory Exercise; Colony Hybridization with an egfp DNA Probe: Part 2; Lab Session 9; Characterization of Recombinant Clones; Lab Session 9A; Completion of Colony Hybridization with mAB Probe; Introduction
- Laboratory Exercise Colony Hybridization with an a-GFP monoclonal Antibody Probe: Part 2; Lab Session 9B; PCR Screening; Introduction; Polymerase Chain Reaction; Laboratory Exercise; Polymerase Chain Reaction Screen for Recombinant Clones: Part 1; Lab Session 9C; Visualization of Green Fluorescent Protein: Part 1; Introduction; Laboratory Exercise; Green Fluorescence Assay and Preparation of a Fresh Master Plate; Lab Session 10; Further Characterization of Recombinant Clones; Lab Session 10A:; Interim Laboratory Session; Laboratory Exercise; Inoculate Cultures for Minipreps; Lab Session 10B