QIAGEN Plasmid Plus Mega Kit (5)

From QIAGEN, this large-scale plasmid preparation kit provides an ultrafast vacuum- and centrifuge-based workflow for purifying transfection-grade plasmid DNA. The Mega-scale procedure can be completed in 40 minutes, and the vacuum manifold design supports parallel processing of up to 24 samples (12 for the Mega/Giga format). The kit produces highly concentrated DNA, typically >1 µg/µl, with endotoxin levels usually below 1 EU/µg, ready for immediate use in applications such as enzymatic modification, cloning, in vitro transcription, in vitro translation, sequencing, and transfection into sensitive cell lines.

The workflow uses QIAfilter Mega-Giga cartridges for rapid lysate clearing and QIAGEN Plasmid Plus spin columns assembled on a vacuum manifold such as the QIAvac 24 Plus. For the Mega format, the maximum recommended LB culture volume is 500 ml with a 12–16 hour incubation. The optional LyseBlue reagent provides visual confirmation of optimum cell lysis and neutralization, and optimized high-yield protocols with extra buffer volumes are provided. Yields reach up to 2.5 mg for the Mega format, within an overall range of 250 µg (Midi) to 10 mg (Giga) across the kit family.

The kit can be stored at 15–25°C for up to 24 months if not otherwise stated on the label. Per the manufacturer's protocol, harvested bacterial culture is centrifuged at 6000 × g for 15 min at 4°C, and after binding, washing and membrane drying, DNA is eluted in 1 ml Buffer EB by centrifugation at 5000 × g for 5 min.