Molecular Cloning: A Laboratory Manual, Book 3 |
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Results 1-3 of 69
Page 1652
... 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 1660
... 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. To obtain high levels of transcription from the phoA promoter , grow an ... C in a microfuge to pellet the cells . Discard the supernatant . b . Suspend ... minutes . Lysozyme will not work efficiently if the pH of the solution is less ...
... 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 . Suspend ... minutes . Lysozyme will not work efficiently if the pH of the solution is less ...
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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