Molecular Cloning: A Laboratory Manual, Book 3 |
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Results 1-3 of 29
Page 17-32
... pellet the cells . Discard the supernatant . • b . Suspend the cell pellets in 100 μl of a freshly prepared solution of lysozyme ( 1 mg / ml ) , 20 % w / v sucrose , 30 mM Tris Cl ( pH 8.0 ) , 1 mM EDTA ( pH 8.0 ) , and place on ice for ...
... pellet the cells . Discard the supernatant . • b . Suspend the cell pellets in 100 μl of a freshly prepared solution of lysozyme ( 1 mg / ml ) , 20 % w / v sucrose , 30 mM Tris Cl ( pH 8.0 ) , 1 mM EDTA ( pH 8.0 ) , and place on ice for ...
Page 17-40
... pellet in 100 μl of H2O . 7. Remove 10 - μl samples of each supernatant and each resuspended pellet . Mix each with 10 μl of 2 × SDS gel - loading buffer and analyze by SDS- polyacrylamide gel electrophoresis to determine the ...
... pellet in 100 μl of H2O . 7. Remove 10 - μl samples of each supernatant and each resuspended pellet . Mix each with 10 μl of 2 × SDS gel - loading buffer and analyze by SDS- polyacrylamide gel electrophoresis to determine the ...
Page 18-40
... pellet as dry as possible . Take care to remove any beads of fluid that adhere to the sides of the microfuge tube . 2 4. Resuspend the pellet by vortexing in 25 μl of H2O . As soon as the pellet is dispersed , add 25 μl of 2 × SDS gel ...
... pellet as dry as possible . Take care to remove any beads of fluid that adhere to the sides of the microfuge tube . 2 4. Resuspend the pellet by vortexing in 25 μl of H2O . As soon as the pellet is dispersed , add 25 μl of 2 × SDS gel ...
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Common terms and phrases
acetate acrylamide activity Adjust the volume agarose aliquots amino acids amount antibody antigen aqueous phase assay autoclaving autoradiographic bacteriophage T4 DNA Biol blunt-ended bromophenol blue cDNA centrifugation chromatography cloning column containing deionized H₂O Dissolve dithiothreitol DNA ligase DNA polymerase dNTPs double-stranded EDTA EDTA pH 8.0 efficiency eluted equal volume Escherichia coli ethanol ethidium bromide expression extract final concentration gene H H OH Incubate intensifying screen lb/sq ligation linkers lysate mammalian cells medium method mg/ml microfuge tube mixture mRNAs NaCl Natl nitrocellulose nitrocellulose filter nucleic acid nucleotides OH OH P.O. Box pellet phenol phosphate pipette plasmid PMSF precipitation prepared protein radioactive radiolabeled reaction remove restriction enzyme room temperature sample Saran Wrap SDS-polyacrylamide gel Sephadex sequences single-stranded sodium sterile stock solution strain supernatant syringe target protein Telephone termini transfected transfer Tris Cl pH vectors wash western blotting µg/ml