Molecular Cloning: A Laboratory Manual, Book 3 |
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Results 1-3 of 38
Page 17-32
... supernatant . b . Suspend the cell pellets in 100 μl of a freshly prepared solution of lysozyme ( 1 mg / ml ) , 20 % w / v sucrose , 30 mм Tris · Cl ( pH 8.0 ) , 1 mм EDTA ( pH 8.0 ) , and place on ice for 10 minutes . Lysozyme will not ...
... supernatant . b . Suspend the cell pellets in 100 μl of a freshly prepared solution of lysozyme ( 1 mg / ml ) , 20 % w / v sucrose , 30 mм Tris · Cl ( pH 8.0 ) , 1 mм EDTA ( pH 8.0 ) , and place on ice for 10 minutes . Lysozyme will not ...
Page 18-15
... supernatant , and resuspend the pellet in acetone ( -20 ° C ) using the same amount that was used in step 2. Vigorous vortexing may be necessary to achieve complete resuspension . Store the suspension on ice for 10 minutes . 4 ...
... supernatant , and resuspend the pellet in acetone ( -20 ° C ) using the same amount that was used in step 2. Vigorous vortexing may be necessary to achieve complete resuspension . Store the suspension on ice for 10 minutes . 4 ...
Page 18-35
... supernatant ( unless the protein of interest is a secretory protein ) . · 2. Resuspend the cells by vortexing in 1 ml of ice - cold 50 mM Tris Cl ( pH 8.0 ) . Recover the cells by centrifugation as described in step 1. Pour off the ...
... supernatant ( unless the protein of interest is a secretory protein ) . · 2. Resuspend the cells by vortexing in 1 ml of ice - cold 50 mM Tris Cl ( pH 8.0 ) . Recover the cells by centrifugation as described in step 1. Pour off the ...
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Common terms and phrases
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