PureLink™ Pro Quick96 Plasmid Purification Kit (Invitrogen™)

The PureLink™ Pro Quick96 Plasmid Kit combines advanced silica plate extraction chemistry with an optimized 96-well plate design for manual or automated processing of plasmid DNA from E. coli in as little as 45 minutes (Table 1). Using the PureLink™ Pro Quick96 Plasmid Kit, you will:

• Obtain consistently high yields of high-purity plasmid DNA
• Experience ease of use and better performance due to improved plate design and protocol
• Increase processing flexibility with protocols for centrifuge, vacuum manifold, or automated platforms

How it works
The PureLink™ Pro Quick96 Plasmid Kit uses two 96-well plates, a filtration plate (clear O-ring), and a binding plate (red O-ring) to purify plasmid DNA. Bacterial cells are subjected to alkaline lysis and are pelleted by centrifugation. The bacterial lysate is then applied to the Quick96 Filtration Plate for clearing and collection in the Quick96 Binding Plate, where the plasmid DNA binds to the silica membrane. After washing steps to remove contaminants, purified DNA is eluted and ready for use in downstream applications. The entire procedure can be completed in 45 minutes (Figure 1).

Improved plate design
The PureLink™ Pro Quick96 Plasmid Kit provides optimized semiskirted 96-well plates for improved performance and results (Figure 2). The Quick96 Filtration Plate is designed for fast and efficient clearing of the bacterial lysates. The Quick96 Binding Plate offers an increased binding capacity of up to 20 μg plasmid DNA. In addition, the plate nozzles with annular collar prevent cross-contamination of samples and improve drying of the silica membrane to prevent ethanol carryover. In addition, sample processing can be performed manually using a benchtop vacuum manifold, by centrifugation, or with an automated liquid handling platform. The semiskirted plate design is compatible with most vacuum manifolds on robotic workstations.

Higher yields and purity
The PureLink™ Pro Quick96 Plasmid Kit allows rapid and reproducible purification of up to 20 μg of plasmid DNA from E. coli strains grown in up to 5 mL LB, 3 mL 2xYT, or 3 mL TB. In an automated platform using 1.5 mL E. coli culture grown overnight in a square-well block, typical yields are 5-15 μg plasmid DNA, with an average of 9.4 μg (Figure 3). The resulting plasmid DNA is supercoiled with no detectable genomic DNA or RNA (Figure 4). The PureLink™ Pro Quick96 Plasmid Kit provides higher yields of high-quality plasmid DNA for use in common downstream applications such as automated sequencing, PCR, restriction enzyme digestion, and cloning.