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SV40 DNA transfection of cells in suspension: analysis of efficiency of transcription and translation of T-antigen |
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| Authors: | G Chu P A Sharp |
| Affiliation: | Center for Cancer Research and Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139 U.S.A. |
| Abstract: | A modification of the Graham and Van der Eb (1974) DNA-calcium phosphate coprecipitation technique is shown to routinely transfect 15% of CV-1 cells with SV40 DNA. The transfection is done in suspension after detachment of cells by trypsin digestion. Transfection efficiency was measured by staining cells for the presence of SV40 T-antigen by indirect immunofluorescence and by assaying for the presence of SV40 early message by the Berk and Sharp (1978) technique. |
| Keywords: | DNA-calcium phosphate coprecipitation immunofluorescence assay early mRNA cloned DNA gel electrophoresis restriction fragments recombinant DNA bp base pairs DME Dulbecco's modified Eagle's medium HeBS Hepes-buffered saline m.o.i. multiplicity of infection PFU plaque-forming units SV40 simian virus 40 |
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