Molecular Cloning: A Laboratory Manual, Book 3 |
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Page 35
assaying for transient expression of the transfected DNA or replating the cells in
the appropriate selective medium for the isolation of stable transformants . b . In
many instances , uptake of DNA is increased by concurrent treatment of the cells
...
assaying for transient expression of the transfected DNA or replating the cells in
the appropriate selective medium for the isolation of stable transformants . b . In
many instances , uptake of DNA is increased by concurrent treatment of the cells
...
Page 47
Harvest exponentially growing CHO cells by trypsinization , and replate them at a
density of 5 x 10 cells per 90 - mm tissue culture dish in 10 ml of alpha medium (
GIBCO ) containing 10 % fetal calf serum . Incubate the cultures for 18 – 20 ...
Harvest exponentially growing CHO cells by trypsinization , and replate them at a
density of 5 x 10 cells per 90 - mm tissue culture dish in 10 ml of alpha medium (
GIBCO ) containing 10 % fetal calf serum . Incubate the cultures for 18 – 20 ...
Page 28
90 Volume of medium Size of petri dish or well ( mm ) 1 . 0 ml 250 ul 60 100 ul 35
100 ul 30 ( well ) The plates should be rocked every 15 minutes to ensure that the
cells do not dry out . When radiolabeling cells for extended periods of time ...
90 Volume of medium Size of petri dish or well ( mm ) 1 . 0 ml 250 ul 60 100 ul 35
100 ul 30 ( well ) The plates should be rocked every 15 minutes to ensure that the
cells do not dry out . When radiolabeling cells for extended periods of time ...
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Contents
BOOK | 3 |
Identification and Analysis of Recombinants 2 108 | 30 |
TRANSFECTION MEDIATED BY DEAEDEXTRAN 16 | 41 |
Copyright | |
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Common terms and phrases
acetate activity added Adjust aliquots allow amount antibody antigen Appendix appropriate approximately aqueous assay bacterial bacteriophage base binding buffer carried cells centrifugation Chapter chromatography Cl pH cloned coli column complete concentration containing culture deionized water described desired detected dilution Dissolve EDTA efficiency electrophoresis enzyme et al ethanol expression extract Figure filter final fragment gene growth hybridization Incubate levels ligation mammalian cells medium method mg/ml microfuge tube minutes mixture mRNAs nucleic acid pellet phase phosphate Place plasmid plate polymerase precipitation prepared procedure Products promoter protein radioactive radiolabeled reaction remove restriction room temperature sample screen selection sequence sodium solution specific step sterile stored strain TABLE Telephone termini tion transcription transfection transfer transformation Tris ug/ml usually vectors volume wash western blotting ОН