Molecular Cloning: A Laboratory Manual, Book 3 |
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Page 16-58
... prepared for measurement of CAT activity . An enzyme frequently used for this purpose is E. coli B - galactosidase , which is expressed in transfected mammalian cells from a promoter with a broad host range ( the SV40 early promoter or ...
... prepared for measurement of CAT activity . An enzyme frequently used for this purpose is E. coli B - galactosidase , which is expressed in transfected mammalian cells from a promoter with a broad host range ( the SV40 early promoter or ...
Page 16-63
... prepared as described on pages 16.59 and 16.60 , steps 1-3 ( except reserve 30 μl of the extract for the assay ) , are assayed as follows : 1. Prepare CAT reaction mixture 2 : 8 mm chloramphenicol cell extract 20 μ . 30 μ . 0.25 M Tris ...
... prepared as described on pages 16.59 and 16.60 , steps 1-3 ( except reserve 30 μl of the extract for the assay ) , are assayed as follows : 1. Prepare CAT reaction mixture 2 : 8 mm chloramphenicol cell extract 20 μ . 30 μ . 0.25 M Tris ...
Page 17-25
... prepared by one of the methods described above ( see pages 17.18-17.24 ) . In most cases , a blunt - ended fragment will be inserted and this step will not be required . In a few cases , the gene fragment prepared as described on pages ...
... prepared by one of the methods described above ( see pages 17.18-17.24 ) . In most cases , a blunt - ended fragment will be inserted and this step will not be required . In a few cases , the gene fragment prepared as described on pages ...
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