Calcium chloride plasmid transfection - (Jun/18/2012 )
I am doing a calcium chloride transfection, but I accidentally put too much calcium chloride into the master ! Instead of water, I put in calcium chloride, and added my plasmids at the needed strength. What I am wondering is, will storing the plasmids in the solution of calcium chloride be detrimental to them? They are lentiviral plasmids around 5kb. The solution is made of:
2M calcium chloride, 1.108 mL
About 42 micrograms of plasmid DNA, 73 microliters
Any advice? Are my plasmids okay? Can I save them? Can I use the calcium chloride plus plasmids in future transfections? I wasted a lot of plasmids in the mix and I hate to think of making another maxiprep...
You can probably precipitate the DNA by adding 2.5 volumes of ethanol and spinning it down. Wash with 70% ethanol a few times and resuspend.