Molecular Cloning: A Laboratory Manual, Book 3 |
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Page 16-47
... containing 10 % fetal calf serum . Incubate the cultures for 18-20 hours at 37 ° C in a humidified incubator in an atmo- sphere of 5-7 % CO2 . 2. Replace the medium with 3 ml of serum - containing medium containing DNA ( 5 ng to 40 μg ...
... containing 10 % fetal calf serum . Incubate the cultures for 18-20 hours at 37 ° C in a humidified incubator in an atmo- sphere of 5-7 % CO2 . 2. Replace the medium with 3 ml of serum - containing medium containing DNA ( 5 ng to 40 μg ...
Page 17-8
... containing ampicillin ( 100 μg / ml ) . Incubate overnight at 37 ° C ( or at 30 ° C for PEX ) . b . Inoculate 5 ml of LB medium containing ampicillin ( 100 μg / ml ) with 50 μl of each overnight culture . Incubate for 2 hours at 37 ° C ...
... containing ampicillin ( 100 μg / ml ) . Incubate overnight at 37 ° C ( or at 30 ° C for PEX ) . b . Inoculate 5 ml of LB medium containing ampicillin ( 100 μg / ml ) with 50 μl of each overnight culture . Incubate for 2 hours at 37 ° C ...
Page B-15
... containing 0.5 % SDS and 100 μg / ml denatured , fragmented salmon sperm DNA ) . 50 × Denhardt's reagent contains 5 g of Ficoll ( Type 400 , Pharmacia ) , 5 g of polyvinylpyrrolidone , 5 g of bovine serum albumin ( Fraction V ; Sigma ) ...
... containing 0.5 % SDS and 100 μg / ml denatured , fragmented salmon sperm DNA ) . 50 × Denhardt's reagent contains 5 g of Ficoll ( Type 400 , Pharmacia ) , 5 g of polyvinylpyrrolidone , 5 g of bovine serum albumin ( Fraction V ; Sigma ) ...
Contents
Selectable Markers 1 | 16-7 |
VECTOR SYSTEMS 16 | 16-17 |
DEVELOPMENT OF PLASMID CLONING VECTORS | 16-24 |
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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