midiprep plasmids not expressing
Posted 20 March 2011 - 10:46 PM
Posted 21 March 2011 - 09:17 AM
If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.
Posted 21 March 2011 - 09:56 AM
Never ever trust an spectrophotometer when dealing with DNA (1st law of molecular biology!) ...only the agarose gel tells you the truth!
Maybe it is just genomic DNA you purified ...who knows ...or just sheared plasmid DNA that you broke in 10000000 pieces?
I would not dry the DNA overnight ...thats just to much ...15 mins should be more than enough! Overdried DNA is hard to solubilize. #
And if you don't feel comfortable with the Qiagen Midi (i don't like em either) ...try another one (e.g. for transfection grade plasmids the Machery&Nagel NucleoBond Xtra Midi EF with the Finializer is good choice!)