Cell-colony hybridization on a nitrocellulose filter to identify clones expressing a transfected gene.
Cells that are transfected with a growth-inhibitory gene can be isolated by using a dominant selective marker. Such isolated clones, however, do not always express the transfected gene. Therefore, clones in which a transfected gene is integrated and expressed should be selected further. We have developed a simple method of identifying such expression-positive clones. We modified the filter hybridization process so that cultured cells are directly transferred to a nitrocellulose filter, immobilized, and hybridized with radiolabeled DNA probes. This method is sensitive enough to detect the presence or the expression of a single-copy gene, and ideal for identifying clones that stably express a transfected gene.
Takahashi
R
R
Department of Pathology and Tumor Biology, Faculty of Medicine, Kyoto University, Japan.
Ariyasu
T
T
Maeda
S
S
Inagaki
T
T
Sugiyama
T
T
D002478:Cells, Cultured; D002999:Clone Cells; D003101:Collodion; D015342:DNA Probes; D005374:Filtration; D006801:Humans; D009693:Nucleic Acid Hybridization; D014162:Transfection
D015342:DNA Probes; D003101:Collodion