Molecular Cloning: A Laboratory Manual, Book 3 |
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Page 16-34
... mixture for 20-30 minutes at room temperature , during which time a fine precipitate should form . At the end of the incubation , pipette the mixture up and down once to resuspend the precipitate . A commonly used alternative procedure ...
... mixture for 20-30 minutes at room temperature , during which time a fine precipitate should form . At the end of the incubation , pipette the mixture up and down once to resuspend the precipitate . A commonly used alternative procedure ...
Page 16-63
... mixture are : 2.3 mM chloramphenicol , 0.11 M Tris Cl , and 129 μM ( 1 μCi / ml ) 1C - labeled acetyl coenzyme A. For each 30 μl of cell extract to be tested , 70 μl of CAT reaction mixture 2 is required . 2. Add 30 μl of cell extract ...
... mixture are : 2.3 mM chloramphenicol , 0.11 M Tris Cl , and 129 μM ( 1 μCi / ml ) 1C - labeled acetyl coenzyme A. For each 30 μl of cell extract to be tested , 70 μl of CAT reaction mixture 2 is required . 2. Add 30 μl of cell extract ...
Page 16-64
... mixture for 15 minutes at 65 ° C to inactivate deacetylases in the extract . Transfer the vial to a water bath set at 37 ° C . 2. Add 200 μl of freshly prepared CAT reaction mixture 3 : 1 м Tris . Cl ( pH 7.8 ) 5 mm chloramphenicol " H ...
... mixture for 15 minutes at 65 ° C to inactivate deacetylases in the extract . Transfer the vial to a water bath set at 37 ° C . 2. Add 200 μl of freshly prepared CAT reaction mixture 3 : 1 м Tris . Cl ( pH 7.8 ) 5 mm chloramphenicol " H ...
Contents
Selectable Markers 1 | 16-7 |
VECTOR SYSTEMS 16 | 16-17 |
DEVELOPMENT OF PLASMID CLONING VECTORS | 16-24 |
Copyright | |
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Acad acetate acrylamide activity Adjust the volume agarose aliquots amount antibody antigen antisera aqueous assay autoclaving bacterial bacteriophage T4 BamHI Biol calcium cDNA cell lines centrifugation chloramphenicol cloned gene column containing culture deionized H₂O dhfr dilution Dissolve dithiothreitol DNA polymerase EDTA EDTA pH 8.0 efficiency elution encoding Escherichia coli ethanol ethidium bromide eukaryotic extract fragment glycerol hybridization immunoglobulin immunoprecipitation Incubate ligation lysate mammalian cells medium method mg/ml microfuge tube minutes at 4°C minutes at room mixture mRNAs NaCl Natl nitrocellulose nitrocellulose filter nucleic acid pellet phosphate plasmid plate PMSF polyacrylamide polyclonal precipitate prepared promoter protoplasts radioactive radiolabeled reaction reagent recombinant remove restriction enzyme Resuspend room temperature sample SDS gel-loading buffer SDS-polyacrylamide gel sequences serum sodium sterile stock solution stored strain supernatant target protein Telephone termini transcription transfection transfer transient expression Tris Cl pH wash western blotting µg/ml