Qiagen
QIAGEN Plasmid Maxi Kit
12165
Info:Supplier Page
We work a lot with molecular biology in our lab, engineering recombinant viruses and cloning into bacteria. To that end, we need high-quality plasmid DNA and also a large volume of it. This kit is used daily for this purpose. Since we often work with low-copy plasmids, we prepare a larger inoculum (usually 300-400 mL) and proceed as described by the supplier's protocol. I have made a few modifications on my own to improve DNA yield, such as pre-warming water or elution buffer before adding to the column and also incubating the column for ~3 min before eluting DNA. This helps get more DNA out of the silica. Also, the protocol suggests eluting in 500 uL of water/elution buffer, instead, I elute in 150 uL (first elution with 100 uL and a second one with 50 uL). This helped a lot in terms of getting a highly concentrated sample.
Plasmid DNA extraction
Bacteria culture
Pre-warm water/elution buffer to 50 °C; Incubate column for 3-5 min before elution; Reduce water/elution buffer volume to increase DNA concentration.
Recombinant plasmid DNA was extracted from 300 mL inoculum (E. coli TOP10, LB media supplemented with ampicillin, 16 h incubation). Concentration was around 1500 ng/uL and 260/280 ratio indicated high quality, contaminant-free DNA sample.
N/A
Fast protocol, and high quality DNA.
Nothing.
Overall, I would definitely recommend this kit for plasmid extraction of a large batch of bacteria culture. It works great and gives high quality DNA for any downstream applications.