Molecular Cloning: A Laboratory Manual, Book 3 |
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Page 16-38
A Laboratory Manual Joseph Sambrook, Tom Maniatis. Calcium Phosphate - mediated Transfection of Cells Growing in Suspension 1. Prepare the solutions and form the calcium phosphate - DNA coprecipitate as described on pages 16.33-16.34 ...
A Laboratory Manual Joseph Sambrook, Tom Maniatis. Calcium Phosphate - mediated Transfection of Cells Growing in Suspension 1. Prepare the solutions and form the calcium phosphate - DNA coprecipitate as described on pages 16.33-16.34 ...
Page E-30
... phosphate groups of the polynucleotide backbone and calcium residues in the resin . Bound nucleic acids can be eluted in phosphate buffers at elevated temperature ( 60 ° C ) . Single - stranded nucleic acids bind less tightly to hy ...
... phosphate groups of the polynucleotide backbone and calcium residues in the resin . Bound nucleic acids can be eluted in phosphate buffers at elevated temperature ( 60 ° C ) . Single - stranded nucleic acids bind less tightly to hy ...
Page E-32
... phosphate ( pH 6.8 ) , whereas doube - stranded DNA is not removed from the column until the phosphate concentration exceeds 0.36 M. 2. Prepare SS and DS buffers by diluting 2 м sodium phosphate ( pH 6.8 ) . 2 M sodium phosphate ( pH ...
... phosphate ( pH 6.8 ) , whereas doube - stranded DNA is not removed from the column until the phosphate concentration exceeds 0.36 M. 2. Prepare SS and DS buffers by diluting 2 м sodium phosphate ( pH 6.8 ) . 2 M sodium phosphate ( pH ...
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