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96 Well Plasmid Kit (96PDV04/96PDV10)

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 40 minutes. The purified plasmid DNA is ready for use in restriction enzyme digestion, ligation, PCR, and sequencing reactions.

DOCUMENTS

DESCRIPTION

Specifications (Cat. # 96PDV04, 96PDV10)

Purify 96 wells of pure plasmid DNA in 40 minutes!
Sample Volume: 1-2 ml of cultured bacterial cells (OD₆₀₀=2-3) per well with using Filter Plate/ 1-5 ml of cultured bacterial cells (OD₆₀₀=2-3) per well without using Filter Plate
Yield: 4-6 μg/ml of cultured bacterial cells (high-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

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
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 (OD₆₀₀=2-3). 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.

Presto™ 96 Well Plasmid Kit Interactive Protocol

Presto™ 96 Well Plasmid Kit Sequencing Data

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.

Plasmid DNA Kit Specifications

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 without filter plate 1-2 ml with filter plate
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	4-6 µg/ml bacteria culture
Elution Volume	30-100 µl	30-100 µl	50-100 µl	100 µl
Operation Time	<15 minutes	<15 minutes	<15 minutes	<60 minutes

RELATED PRODUCTS

Product Name	RXNS	Catalogue Number
Presto™ Mini Plasmid Kit	100, 300	PDH100/300
High-Speed Plasmid Mini Kit	100, 300	PD100/300
High-Speed Plasmid Mini Kit (10-50 Kb)	100, 300	PDL100/300
High-Speed Plasmid Advance Kit (50-100 ml)	25	PA025
Presto™ Midi Plasmid Kit	25	PIF025
Presto™ Midi Column	25	PIF01
Presto™ Midi Plasmid Kit (Endotoxin Free)	25	PIFE25
Geneaid™ Midi Plasmid Kit	25	PI025
Geneaid™ Midi Plasmid Kit (Endotoxin Free)	25	PIE25
Presto™ Plasmid DNA Concentration Kit	25, 50, 100	PC0250/500/1000
Geneaid™ Maxi Plasmid Kit	10, 25	PM010/25
Geneaid™ Maxi Plasmid Kit (Endotoxin Free)	10, 25	PME10/25
Presto™ 96 Well Plasmid Kit	4 x 96, 10 x 96	96PDC04/10 96PDV04/10

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