Modified miniprep method for the rapid recovery of episomes from transfected breast epithelial cells

Episomal vectors such as pCEP4 ara useful in expression cloning because they can replicate in both prokaryotes and eukaryotic cells. We have found a rapid ann efficient means of extracting them from transfected MCF-10A nonmalignant human breast epithelial cells. Ne show that a plasmid miniprep protocol, modified by the addition of an extraction that eliminates a DNase activity can consistently harvest pCEP4 episomes from the transfected cells (516 +/- 112 pg/harvest, mean +/- standard deviation; n = II). The quality of the episomal DNA obtained in this manner,was verified by PCR, Southern blot and the retransformation of Escherichia coli. This simple method enables the efficient recovery! of episomes and is applicable in the expression cloning of potential oncogenes using host MCF-10A cells.