Molecular Cloning: A Laboratory Manual, Book 3 |
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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 17-41
... supernatant and set aside for the next step . Resuspend the pellet in 100 μl of 1 × SDS gel - loading buffer . 7. Remove 10 - μl samples of the supernatant and resuspended pellet . Mix the supernatant sample with 10 μl of 2 × SDS gel ...
... supernatant and set aside for the next step . Resuspend the pellet in 100 μl of 1 × SDS gel - loading buffer . 7. Remove 10 - μl samples of the supernatant and resuspended pellet . Mix the supernatant sample with 10 μl of 2 × SDS gel ...
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 mм 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 mм 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
acetate acrylamide Adjust the volume agarose aliquots amino acids amount antibody antigen aqueous phase assay autoclaving autoradiographic bacteriophage T4 DNA blunt-ended bromophenol blue cDNA centrifugation chromatography cloning column containing deionized H₂O Dissolve dithiothreitol DNA polymerase dNTPs double-stranded EDTA EDTA pH 8.0 efficiency eluted Escherichia coli ethanol ethidium bromide expression extract final concentration fragment gene H H H H OH hybridization Incubate intensifying screen lb/sq ligation linkers lysate mammalian cells medium method mg/ml microfuge tube mixture Molecular mRNAs NaCl Natl NH3 H nitrocellulose nitrocellulose filter nucleic acid nucleotides OH OH P.O. Box pasteur pipette pellet phenol phosphate pipette plasmid PMSF precipitation prepared protein radioactive radiolabeled reaction remove restriction enzyme room temperature sample Saran Wrap SDS-polyacrylamide gel sequence single-stranded sodium sterile stock solution strain supernatant syringe target protein Telephone termini transfected transfer Tris Cl pH vectors wash western blotting µg/ml