pCMVβ DNA was prepared using the indicated preparation method. Huh-7 cells (4 x 104) were transfected using 200 ng plasmid DNA and 0.75 μl Attractene Transfection Reagent. β-gal activity and protein content were measured after 48 hours. Plasmid DNA prepared with QIAGEN Plasmid Plus 96 Miniprep and BioRobot Kits resulted in highly efficient transfection into sensitive cell lines.
Using the QIAGEN Plasmid Plus 96 Miniprep Kit, pCMVβ plasmid DNA was purified from E. coli DH5a cultures grown in LB medium. DNA was quantified using absorbance measurement and verified by agarose gel analysis. High yields were achieved with the QIAGEN Plasmid Plus 96 Miniprep Kit.
pCMVβ plasmid DNA was purified from 1.3 ml and 5 ml E. coli DH10B cultures using the QIAGEN Plasmid Plus 96 Miniprep Kit, the QIAwell 96 Ultra Plasmid Kit, and kits from other suppliers. Endotoxin levels were determined using the Kinetic-QCL Kinetic Chromogenic Assay. For the EndoFree Plasmid Giga Kit, the endotoxin content was 0.0005 EU/μg DNA. EF: EndoFree Plasmid Giga Kit. QPP96: QIAGEN Plasmid Plus 96 Miniprep Kit. QW96: QIAwell 96 Ultra Plasmid Kit.