The osmolarity of the electroporation medium affects the transient expression of genes
During transfection on 5123 and FT02B hepatoma cells and NIH 3T3 fibroblast cells by electroporation, it was observed that the expression of the reporter gene and the viability of the cells was very sensitive to varying amounts of DNA solution added, despite a constant setting of voltage and capacit...
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| Veröffentlicht in: | Nucleic acids research 1990-11, Vol.18 (21), p.6464-6464 |
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| container_title | Nucleic acids research |
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| creator | VAN DEN HOFF, M. J. B LABRUYERE, W. T MOORMAN, A. F. M LAMERS, W. H |
| description | During transfection on 5123 and FT02B hepatoma cells and NIH 3T3 fibroblast cells by electroporation, it was observed that the expression of the reporter gene and the viability of the cells was very sensitive to varying amounts of DNA solution added, despite a constant setting of voltage and capacity during electroporation. As few quantitative data are available that detail the extent to which the efficiency of electroporation is dependent on medium osmolarity, the following experiment was performed according to Potter et al., except that the complete procedure was done at room temperature which resulted in an increase of the specific activity of CAT by approximately 25%. The data clearly indicate that the use of hypotonic medium during electroporation has to be avoided. |
| doi_str_mv | 10.1093/nar/18.21.6464 |
| format | Article |
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J. B</creatorcontrib><creatorcontrib>LABRUYERE, W. T</creatorcontrib><creatorcontrib>MOORMAN, A. F. M</creatorcontrib><creatorcontrib>LAMERS, W. H</creatorcontrib><title>The osmolarity of the electroporation medium affects the transient expression of genes</title><title>Nucleic acids research</title><addtitle>Nucleic Acids Res</addtitle><description>During transfection on 5123 and FT02B hepatoma cells and NIH 3T3 fibroblast cells by electroporation, it was observed that the expression of the reporter gene and the viability of the cells was very sensitive to varying amounts of DNA solution added, despite a constant setting of voltage and capacity during electroporation. As few quantitative data are available that detail the extent to which the efficiency of electroporation is dependent on medium osmolarity, the following experiment was performed according to Potter et al., except that the complete procedure was done at room temperature which resulted in an increase of the specific activity of CAT by approximately 25%. The data clearly indicate that the use of hypotonic medium during electroporation has to be avoided.</description><subject>Animals</subject><subject>Avian Sarcoma Viruses - genetics</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cell Line</subject><subject>Cell Survival</subject><subject>Chloramphenicol O-Acetyltransferase - biosynthesis</subject><subject>Chloramphenicol O-Acetyltransferase - genetics</subject><subject>Electric Stimulation - methods</subject><subject>electroporation</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>Genetic Techniques</subject><subject>Methods. Procedures. 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H</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c476t-7c5110c10e8ed7e7c18b776e895313d7ff13d09baa75132f63a51222315d4813</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1990</creationdate><topic>Animals</topic><topic>Avian Sarcoma Viruses - genetics</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Cell Line</topic><topic>Cell Survival</topic><topic>Chloramphenicol O-Acetyltransferase - biosynthesis</topic><topic>Chloramphenicol O-Acetyltransferase - genetics</topic><topic>Electric Stimulation - methods</topic><topic>electroporation</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>Genetic Techniques</topic><topic>Methods. Procedures. 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| identifier | ISSN: 0305-1048 |
| ispartof | Nucleic acids research, 1990-11, Vol.18 (21), p.6464-6464 |
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| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_332585 |
| source | MEDLINE; PubMed Central; Oxford University Press Journals Digital Archive Legacy |
| subjects | Animals Avian Sarcoma Viruses - genetics Biological and medical sciences Biotechnology Cell Line Cell Survival Chloramphenicol O-Acetyltransferase - biosynthesis Chloramphenicol O-Acetyltransferase - genetics Electric Stimulation - methods electroporation Fundamental and applied biological sciences. Psychology Gene Expression Genetic engineering Genetic technics Genetic Techniques Methods. Procedures. Technologies Mice Osmolar Concentration Promoter Regions, Genetic Recombinant Fusion Proteins - biosynthesis Transfection |
| title | The osmolarity of the electroporation medium affects the transient expression of genes |
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