Presto™ 96 Well Plasmid Kit

DNA Extraction/Plasmid DNA Purification

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 16-20 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.

Specifications

Purify 96 wells of pure plasmid DNA in 25 minutes!
Sample Volume: 1-5 ml of cultured bacterial cells per well
Yield: 16-20 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)

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 16 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.

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

Figure 1 .Sequencing data of (pBluescript) plasmid DNA purified using the Presto™ 96 Well Plasmid Kit.

E. coli Culture Volume	260/280	260/230	Yield
1.5 ml	1.90	2.10	6-9 µg
4.0 ml	1.90	2.10	16-20 µg

Table 1 .Yield and quality of plasmid DNA purified using the Presto™ 96 Well Plasmid Kit. 5 µl aliquots of a 60 µl eluate of purified super coiled plasmid DNA from 1.5 ml and 4 ml overnight E. coli (DH5α) culture, containing a 3 kb plasmid pBluescript (OD600 = 3.7 U/ml) were EcoRI digestion and analyzed by electrophoresis on a 0.8% agarose gel.

COMPONENTS

P1, P2, P3 Buffers
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

Specifications	Presto™ Midi Plasmid Kit	High-Speed Plasmid Mini Kit	High-Speed Plasmid Mini Kit 10-50 Kb	Presto™ 96 Well Plasmid Kit
Cat. #	PDH100 PDH300	PD100 PD300	PDL100 PDL300	96PDC04/10 96PDV04/10
Format	Spin Column	Spin Column	Spin Column	96 Well Plate
Binding Capacity	50 µg	30 µg	30 µg	35 µg
Culture Input	1-7 ml	1-5 ml	1-4 ml	1-5 ml
Plasmid Size	1-15 kb	1-15 kb	10-50 kb	1-15 kb
Typical Yield	up to 50 µg	10-30 µg	10-25 µg	20-35 µg/well
Elution Volume	30-100 µl	30-100 µl	50-100 µl	100 µl
Operation Time	<15 minutes	<15 minutes	<15 minutes	25 minutes