Molecular Cloning: A Laboratory Manual, Book 3 |
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Page 1638
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 ...
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Acad acetate acrylamide activity agarose aliquots amount antibody antigen aqueous phase assay autoclaving autoradiographic bacterial bacteriophage T4 DNA BamHI Biol blunt-ended cDNA cell lines centrifugation chloramphenicol chromatography cloned gene coli DNA polymerase column containing culture Dissolve dithiothreitol DNA ligase DNA polymerase dNTPs double-stranded EDTA EDTA pH 8.0 efficiency electrophoresis eluted encoding Escherichia coli ethanol ethidium bromide eukaryotic expression vectors extract gel-loading buffer H₂O hybridization Incubate intensifying screen ligation linkers mammalian cells medium method mg/ml microfuge tube mixture mRNAs NaCl Natl nitrocellulose nitrocellulose filter nucleic acid nucleotides OH OH P.O. Box pellet phosphate pipette plasmid PMSF polycloning precipitation prepared Proc promoter radioactive radiolabeled reaction remove restriction enzyme room temperature sample sequences sodium sterile stock solution stored strain supernatant target protein Telephone termini tion transcription transfection transfer Tris Cl pH vectors washed western blotting µg/ml