The Presto™ 96 Well Plasmid Kit was designed for high-throughput purification of plasmid DNA from 1-5 ml of cultured bacterial cells per well using an efficient 96 well filter plate and 96 well binding plate system. A modified alkaline lysis method and RNase treatment are used to obtain clear cell lysate with minimal genomic DNA and RNA contaminants. Typical yields are 20-35 μg for high-copy number plasmid or 3-10 μg for low-copy number plasmid from 4 ml of cultured bacterial cells. DNA phenol extraction or alcohol precipitation is not required and the entire procedure can be completed within 25 minutes. The purified plasmid DNA is ready for use in restriction enzyme digestion, ligation, PCR, and sequencing reactions.

Advantages (Cat. # 96PDC04, 96PDC10, 96PDV04, 96PDV10)

• Purify 96 wells of pure plasmid DNA in 25 minutes!
• Sample Volume: 1-5 ml of cultured bacterial cells per well 
• Yield: 20-35 μg per well (high-copy plasmid), 3-10 μg per well (low-copy plasmid)
• Binding Capacity: 35 μg per well
• Plasmid Size Range: 1-20 kb
• 96 Well plates for filtration and DNA binding
• Elution Volume: 60 μl from 100 μl elution buffer volume
• Centrifuge or vacuum manifold protocol
• Storage: dry at room temperature (15-25ºC)

Applications

Fluorescent and Radioactive Sequencing, Restriction Enzyme Digestion, Library Screening, Ligation, Transformation and PCR

Components

• PD1, PD2, PD3 Buffers
• W1 Buffer
• Wash Buffer
• Elution Buffer (10 mM Tris-HCl, pH8.5 at 25°C)
• RNase A
• Presto™ Plasmid 96 Well Filter Plates
• Presto™ Plasmid 96 Well Binding Plates
• 96 Deep Well Plates
• 0.35 ml Collection Plates
• Adhesive Film

Quality Control

The quality of the Presto™ 96 Well Plasmid Kit is tested on a lot-to-lot basis by isolating plasmid DNA from a 4 ml overnight E. coli (DH5α) culture containing plasmid pBluescript (A600 > 3.7 U/ml). Following the purification process, a yield of more than 20 μg per well is obtained and the A260/A280 ratio is between 1.8-2.0. The purified plasmid DNA (1 µg) is used in EcoRI digestion, and analyzed by electrophoresis.