Molecular Cloning: A Laboratory Manual, Book 3 |
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Results 1-3 of 59
Page 16-52
... for 5 minutes at 4 ° C . Wash the cells twice in ice - cold phosphate- buffered saline ( PBS ; see Appendix B ) . 2. Add to the mammalian cell pellet the protoplasts prepared in step 6 , page 16.49 . Use a ratio of protoplasts to ...
... for 5 minutes at 4 ° C . Wash the cells twice in ice - cold phosphate- buffered saline ( PBS ; see Appendix B ) . 2. Add to the mammalian cell pellet the protoplasts prepared in step 6 , page 16.49 . Use a ratio of protoplasts to ...
Page 16-60
... minutes at 4 ° C in a microfuge . Transfer the supernatant to a fresh microfuge tube . Reserve 50 μl for the CAT assay , and store the remainder of the extract at -20 ° C . 4. Incubate the 50 - μl aliquot of the extract for 10 minutes ...
... minutes at 4 ° C in a microfuge . Transfer the supernatant to a fresh microfuge tube . Reserve 50 μl for the CAT assay , and store the remainder of the extract at -20 ° C . 4. Incubate the 50 - μl aliquot of the extract for 10 minutes ...
Page 17-32
... 4 4. To obtain high levels of transcription from the phoA promoter , grow an ... C in a microfuge to pellet the cells . Discard the supernatant . • b ... minutes . Lysozyme will not work efficiently if the pH of the solution is less than ...
... 4 4. To obtain high levels of transcription from the phoA promoter , grow an ... C in a microfuge to pellet the cells . Discard the supernatant . • b ... minutes . Lysozyme will not work efficiently if the pH of the solution is less than ...
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Common terms and phrases
Acad acetate acrylamide activity agarose aliquots amino acids amount antibody antigen aqueous phase assay autoclaving autoradiographic bacteriophage T4 DNA blunt-ended bromophenol blue cDNA centrifugation chromatography codon coli DNA polymerase column containing culture Dissolve dithiothreitol DNA ligase DNA polymerase dNTPs double-stranded EDTA EDTA pH 8.0 efficiency eluted Escherichia coli ethanol ethidium bromide expression extract H H OH Incubate intensifying screen ligation linkers lysate mammalian cells medium method mg/ml microfuge tube mixture Molecular mRNAs NaCl Natl nitrocellulose nitrocellulose filter nucleic acid nucleotides OH OH P.O. Box pasteur pipette pellet phenol phosphate pipette plasmid PMSF polyclonal precipitation prepared promoter radioactive radiolabeled reaction remove restriction enzyme room temperature sample Saran Wrap SDS gel-loading buffer SDS-polyacrylamide gel sequence single-stranded sodium sterile stock solution stored strain supernatant syringe target protein Telephone termini transcription transfected transfer Tris Cl pH vectors wash western blotting µg/ml