Purification of DNA for the transfection of a Spodoptera frugiperda cell line.
Spodoptera frugiperda (Sf-9) cells have been widely used in baculovirus expression systems, transient gene expression studies and transgenic cell lines. These applications commonly require the transfection of bacterial plasmid DNA. One of the most reliable methods of preparing transfection-quality plasmid DNA is cesium chloride (CsCl) density gradient centrifugation. However, the traditional CsCl DNA purification is a long and laborious process. We have made a series of modifications to the traditional method that makes it faster, safer and easier. In the current study we demonstrate that DNA prepared by our modified CsCl method was also better for the transfection of Sf-9 cells than DNA prepared by the traditional CsCl method.
Slack
Jeffrey M
JM
United States Department of Agriculture, Agriculture Research Service, Insect Biocontrol Laboratory, Beltsville, MD 20705, USA. lawrence@ba.ars.usda.gov
Lawrence
Susan D
SD
D000818:Animals; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient / Q000295:instrumentation / Q000379:methods*; D002586:Cesium / Q000737:chemistry; D002712:Chlorides / Q000737:chemistry; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D015203:Reproducibility of Results; D018411:Spodoptera; D014162:Transfection / Q000295:instrumentation / Q000379:methods*
D002712:Chlorides; D004269:DNA, Bacterial; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D002586:Cesium; C028019:cesium chloride