Molecular Cloning: A Laboratory Manual, Book 3 |
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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 70
Expression Cloning of Genomic DNA In principle , the genome of a mammalian
species contains all the information necessary for expression of a given gene .
Furthermore , because the cellular machinery for transcription , translation , and ...
Expression Cloning of Genomic DNA In principle , the genome of a mammalian
species contains all the information necessary for expression of a given gene .
Furthermore , because the cellular machinery for transcription , translation , and ...
Page 8
If most of the colonies can be assumed to contain a cDNA ( because directional
cloning or a dephosphorylated vector was used in step 1 ) ... Inoculate 5 ml of LB
medium containing ampicillin ( 100 ug / ml ) with 50 ul of each overnight culture .
If most of the colonies can be assumed to contain a cDNA ( because directional
cloning or a dephosphorylated vector was used in step 1 ) ... Inoculate 5 ml of LB
medium containing ampicillin ( 100 ug / ml ) with 50 ul of each overnight culture .
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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