Qiagen
Plasmid Midi Kit (25)
12143
I used Qiagen's plasmid midi prep kit for large scale extraction of pBAD plasmid DNA from overnight grown Salmonella enterica cultures. Got very good yield of 25 microgram total in 100 microliter volume with quality of 260:280 ratio 1.87.
Large scale plasmid DNA extraction from Salmonella enterica cultures
25 ml of overnight grown Salmonella enterica cultures in LB broth
I used 25 ml of overnight grown S. enterica culture for the extraction. The culture was pelleted by centrifugation at 1000 rpm for 4 minutes and then used for extraction following easy to follow instructions given in the Plasmid Midi kit protocol. Finally eluted in the 100 microliter of nuclease free molecular biology grade water.
I recommend pre-heating nuclease free water to 50 to 55 degree C to elute plasmid DNA. This enhances solubility and recovery or yield.
Final extraction gave me an yield of 25 microgram total plasmid DNA (250 ng/microliter with 260:280 ratio of 1.83).
None
Simple protocol and very easy to perform.
If you need to prep large amount of plasmid DNA and make large stock, then use this plasmid midi kit for plasmid DNA extraction.