Molecular Cloning: A Laboratory Manual, Book 3 |
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Page xii
... DNA Methylation 5.16 Influence of Methylation on DNA Mapping 5.26 DIGESTING DNA WITH RESTRICTION ENZYMES 5.28 ... Ligase 5.62 E. coli DNA Ligase 5.64 Bacteriophage T4 RNA Ligase 5.66 Bacteriophage T4 Polynucleotide Kinase 5.68 Alkaline ...
... DNA Methylation 5.16 Influence of Methylation on DNA Mapping 5.26 DIGESTING DNA WITH RESTRICTION ENZYMES 5.28 ... Ligase 5.62 E. coli DNA Ligase 5.64 Bacteriophage T4 RNA Ligase 5.66 Bacteriophage T4 Polynucleotide Kinase 5.68 Alkaline ...
Page F-7
... DNA ligase mixture is prepared as follows : 1 м Tris Cl ( pH 7.6 ) 100 mM MgCl2 200 mm dithiothreitol 10 mм ATP H2O 1.0 μl 1.0 με 1.0 με 1.0 με 5.5 με bacteriophage T4 DNA ligase 1 Weiss unit Mix the components in a tube stored in an ...
... DNA ligase mixture is prepared as follows : 1 м Tris Cl ( pH 7.6 ) 100 mM MgCl2 200 mm dithiothreitol 10 mм ATP H2O 1.0 μl 1.0 με 1.0 με 1.0 με 5.5 με bacteriophage T4 DNA ligase 1 Weiss unit Mix the components in a tube stored in an ...
Page F-9
... DNA phosphorylated linkers H2O to 15 μl 0.1-0.5 με ( in a volume of 7 μl or less ) 1-2 με ( in a volume of 8 μl or less ) 5 mM ATP 2 μ . 10 blunt - end ligation buffer 2 μ 1 μ bacteriophage T4 DNA ligase ( 1-2 Weiss units ) Incubate the ...
... DNA phosphorylated linkers H2O to 15 μl 0.1-0.5 με ( in a volume of 7 μl or less ) 1-2 με ( in a volume of 8 μl or less ) 5 mM ATP 2 μ . 10 blunt - end ligation buffer 2 μ 1 μ bacteriophage T4 DNA ligase ( 1-2 Weiss units ) Incubate the ...
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