I followed maxiprep protocol and the yield was as low as 5.9ng/ul. All the steps were followed and everything was taken care of. Is it because the cultures were a month old?
You started with a liquid culture right? I use 150 ml of liquid culture and always freshly prepared just one night before the maxiprep. Otherwise, the column of the maxi-prep kit can be expired. However, I faced with the same problem, then I re-performed the maxi-prep. This time everything was perfect. Thus, if you have a chance, try it again.
Tuba Sena Ogurlu Thank You ma'am. I found out that with nuclease free water the yield is better than elution buffer.
Hello Yukti Kataria
For maxiprep your yield should be around 200 – 1000ug.
There are many steps where things can go wrong. But the two major steps could be either the culture or the DNA precipitation step.
I feel the cultures are too old. You should try to extract plasmid DNA from fresh cultures. Extracting plasmid DNA from very old cultures can result in low yield since many bacterial cells are dead and plasmid DNA they contain gets degraded. If you feel that you cannot perform plasmid prep immediately, you may spin down the bacteria and store the pellet in -80°C freezer for later plasmid prep.
Best.
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