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|Tipo de Material:||Recurso Internet|
|Tipo de Documento:||Livro, Recurso Internet|
|Todos os Autores / Contribuintes:||
Dominique Robertson; Scott H Shore; David M Miller
|Descrição:||xix, 204 pages : illustrations ; 28 cm|
|Conteúdos:||Part 1 Manipulation of DNA: Lab session 1 - culturing E. Coli for plasmid isolation: preparation of solid media, making stabs cultures, streaking out plasmid clones on LB/carb plates, growth curve of E. Coli,; cultures for plasmid preparations; Lab session 2 - growth curve of E. Coli: examination of clones, E. Coli growth curve, preparation of solutions for large scale plasmid DNA preps, inoculation of cultures for large scale plasmid DNA preps; Lab session 3A - large scale preparation of plasmid DNA by alkaline lysis; Lab session 3B - purification of plasmid DNA using columns; Lab session 3C - purification of plasmid DNA using cesium gradients; Lab session 3D - purification of plasmid DNA using polyethylene glycol; Lab session 4 - preparation of vector DNA - ethidium bromide spot test, preparation of Hindill Cut, phosphated vector, gel electrophoresis, electrophoresis of pUR288, shrimp alkaline phosphatase treatment of pUR288, overnight cultures for transformation-competent cells; Lab session 5 - transformation of E. Coli - preparation of frozen, transformation-competent cells, test ligations of pUR288 electrophoresis of ligation products, transformation; Lab session 6 - preparation and ligation of insert DNA: analysis of vector preparation, isolation of myo-3 DNA from agarose, ligations; Lab session 7A - transformation of E. Coli with ligated DNA and labelling a DNA probe: CaCl2-mediated transformation, electrophoration, electrophoresis of ligation reactions, random primed DNA labelling with Dig-11-dUTP; Lab session 7B: labelling DNA with radioactive nucleotides: random primer labelling of DNA using 32p-dCTP; Lab Session 8 - DNA probe preparation, replica plating: estimating the yield of the DIG-labelled probe, making replica plates; Lab session 9 - colony hybrid dilations, DNA probe: colony hybridizations - transfer of colonies to nylon membranes, prehybridization, DNA/DNA hybridization, adding DNA probe; Lab session 10 - restriction of map of plasmid p2D, colony nybridizations. Part 2 Restriction enzyme reactions; colony hybridization with a DNA probe - washing filters probed with Dig-11-dUTP labelled myo-3 fragment; colony hybridization with a DNA probe - washing filters probed with 32p-dCTP labelled myo-3 fragment; gel electrophoresis of restriction digests; Lab session 11 - screening colonies with monoclonal antibodies: colony hybridizations - mAb probe, first day, alkaline lysis minipreps, finish restriction maps; Lab session 12 - isolation and characterization of miniprep DNA from potential transformants. (Part contents).|
|Responsabilidade:||Dominique Robertson, Scott Shore, David M. Miller.|
"This is a clearly written and well thought out laboratory manual that covers many of the basic concepts and laboratory techniques common to molecular biology... The book has many clear and concise
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