Minitransfection: a simple, fast technique for transfections.
A fast, simple and inexpensive minitransfection technique, using either a lipofection or a calcium phosphate coprecipitation method to introduce foreign DNA into living cells is presented. This technique is based on the use of 24-well or 96-well tissue culture plates and can be used for both transient and stable transfections. Because it is a microtechnique, only small amounts of DNA, cells and transfection reagents are necessary, and it is easy to handle multiple DNA transfections or cotransfections in different cell lines and in duplicates or triplicates. The technique can be used to study viral gene expression, virus replication and chloramphenicol acetyltransferase (CAT) assay in different cell lines, for example, in the large scale screening and testing of antiviral agents.
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Children's Hospital of Philadelphia, Division of Infectious Diseases and Immunology, Pennsylvania 19104.
Gönczöl
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Srinivasan
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Douglas
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SD
Plotkin
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SA
D000818:Animals; D000911:Antibodies, Monoclonal; D000956:Antigens, Viral / Q000235:genetics; D002130:Calcium Phosphates; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003587:Cytomegalovirus / Q000235:genetics / Q000276:immunology; D004274:DNA, Recombinant / Q000378:metabolism; D015967:Gene Expression Regulation, Viral; D015756:Genes, tat / Q000235:genetics; D016325:HIV Long Terminal Repeat / Q000235:genetics; D006367:HeLa Cells; D017874:Immediate-Early Proteins; D010714:Phosphatidylethanolamines; D010957:Plasmids; D012680:Sensitivity and Specificity; D014162:Transfection; D014407:Tumor Cells, Cultured; D014779:Virus Replication
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D002130:Calcium Phosphates; D004274:DNA, Recombinant; D017874:Immediate-Early Proteins; D010714:Phosphatidylethanolamines; C485828:alpha-tricalcium phosphate; C044582:immediate-early proteins, cytomegalovirus; C485830:tetracalcium phosphate; C485838:calcium phosphate, monobasic, anhydrous; C020888:1,2-dielaidoylphosphatidylethanolamine; C020243:calcium phosphate; D015500:Chloramphenicol O-Acetyltransferase; C485829:calcium phosphate, dibasic, anhydrous