does low plasmid concentration affect transfection rate? - (Dec/24/2007 )
I had a plasmid with 260ug/mL. As transfection process, I have to use 20uL for transfection. Does this affect my transfection rate?
For transfection one has to dilute the DNA in transfection media, so having a few extra microliters, should not be a problem. I havent seen it to be problem. If this is a miniprep DNA, it might not work all the time.
You have to use 20 µl in which volume of transfection reagent + medium for complex formation? e.g. if you would have 20 µl of H2O+DNA in 30 µl total volume, it might be problematic, if you have 20 µl in 200 it won't harm that much. If you have fears: increase concentration of your DNA by precipitation or by vacuüm.
Thanks. I precipitated my products today. WE will see what happens.
the concentration is not really a problem, if not too low (considering less than 50ng/µl should be a pb). Major impact is the purity (see od ratios) and the endotowins. I use a kit to prepare the plasmids for transfection.