Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/04/28
|DNA Concentration of KAH187-MV8 plasmid||Main project page|
Previous entry Next entry
The KAH187-MV8 plasmid is a mammalian expression vector containing the CMV promoter, followed by two copies of AmCyan with a nuclear localization signal. We'll be using it as a control for transfection studies, but the Neon transfection unit asks for plasmid at a concentration of at least 1,000 ng/µL. I earlier performed a plasmid maxiprep and collected >1 mL of KAH187-MV8 at 164 ng/µL. Today I plan to concentrate 10x, for a final concentration of >1,000 ng/µL in 100 µL final volume.
This is a classic nucleic acid precipitation/resuspension protocol. Instructions taken from the Sigma GenElute Plasmid Maxiprep protocol.
Success! With an 88% yield, no less. There's some of the original plasmid left over as well. Both samples stored at -20°C in my 2nd box.