maxiprep from a miniprep - (Sep/28/2006 )
Hi,
I got very low concentrations (around 60ul/ml) for DNA plasmids purified by miniprep. I was wondering if it would be possible to do a maxiprep from the plasmids I got via miniprep:unsure: . And in this case what DNA quantity should I use? (I used normally 10ul of the PCR products and 10 ng of DNA for the postiv control so I though I could use 10 ng as well )
Or maybe I should make a miniprep again (also from the minipreps I got before) and get greater amounts??
Thanks for your help.
retransform ur plasmid. U need only a few picograms of DNA for this. and pick a colony and grow in 100ml of LB or more for maxi prep.
If u have mini prep bacterial culture, i would use 100ul to inoculate 100ml LB media.
do a electroporation with 0.5µl and use a tip for separating baterias on the plate
then use a colony for the maxiprep.
20µl of a miniprep culture are sufficent.