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Scott Witherow 205 $a3rd ed. 210 $aAmsterdam ;$aBoston $cElsevier /Academic Press$dc2012 215 $a1 online resource (xxvi, 200 pages) $ccolor illustrations 300 $aIncludes index. 300 $aPrevious edition: 2006. 311 $a0-12-385544-6 320 $aIncludes bibliographical references and index. 327 $aLab Session 1 Getting Oriented: Practicing with Micropipettes -- Lab Session 2 Purification and Digestion of Plasmid (Vector) DNA -- Lab Session 3 PCR Amplification of egfp and Completion of Vector Preparation -- Lab Session 4 Preparation of Insert DNA (egfp) PCR Product -- Lab Session 5 DNA Ligation and Transformation of Escherichia coli -- Lab Session 6 Colony Hybridization -- Lab Session 7 Characterization of Recombinant Clones: Part 1 -- Lab Session 8 Characterization of Recombinant Clones: Part 2 -- Lab Session 9 Characterization of Recombinant Clones: Part 3 -- Lab Session 10 Computational Analysis of DNA Sequence from a Positive Clone: Part 2 -- Lab Session 11 Expression of Fusion Protein from Positive Clones, SDS-PAGE and Western Blot: Part 1 -- Lab Session 12 Expression of Fusion Protein from Positive Clones, SDS-PAGE and Western Blot: Part 2 -- Lab Session 13 Extraction of Recombinant Protein from Escherichia coli Using a Glutathione Affinity Column -- Lab Session 14 Analysis of Purification Fractions -- Lab Session 15 Total RNA Purification -- Lab Session 16 Analysis of gst::egfp mRNA Levels by RT-qPCR: Part 1 -- Lab Session 17 Analysis of gst::egfp mRNA Levels by RT-qPCR: Part 2 -- Lab Session 18 Analysis of gst::egfp mRNA Levels by Semi-Quantitative RT-PCR: Part 1 -- Lab Session 19 Analysis of gst::egfp mRNA Levels by Semi-Quantitative RT-PCR: Part 2 -- Appendix 1 Equipment -- Appendix 2 Prep List -- Appendix 3 Preparation of Competent E. coli Cells -- Appendix 4 Pre-Lab Questions. 330 $aThis manual is an indispensable tool for introducing advanced undergraduates and beginning graduate students to the techniques of recombinant DNA technology, or gene cloning and expression. 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