Molecular Cloning: A Laboratory Manual, Book 3 |
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Page 46
Depending on the experiment , continue with step 7a ( transient expression ) or
step 7b ( stable transformation ) . 7 . a . Transient expression : Harvest the cells
48 – 60 hours after transfection for analysis of RNA or DNA by hybridization .
Depending on the experiment , continue with step 7a ( transient expression ) or
step 7b ( stable transformation ) . 7 . a . Transient expression : Harvest the cells
48 – 60 hours after transfection for analysis of RNA or DNA by hybridization .
Page 47
The method outlined below works efficiently for stable transformation of CHO
cells by plasmid DNA , yielding approximately 15 - fold more transformants than
calcium phosphate - DNA coprecipitation . However , there is no difference
between ...
The method outlined below works efficiently for stable transformation of CHO
cells by plasmid DNA , yielding approximately 15 - fold more transformants than
calcium phosphate - DNA coprecipitation . However , there is no difference
between ...
Page 52
If the protoplasts lyse at this stage and the solution becomes viscous , the
efficiency of transformation will almost certainly be very low . 3 . Centrifuge the
mixture of bacterial protoplasts and mammalian cells at 2000 rpm for 10 minutes
at room ...
If the protoplasts lyse at this stage and the solution becomes viscous , the
efficiency of transformation will almost certainly be very low . 3 . Centrifuge the
mixture of bacterial protoplasts and mammalian cells at 2000 rpm for 10 minutes
at room ...
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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