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96 Well Plasmid Kit (96PDV04, 96PDV10) (96PDC04, 96PDC10)
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.

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
Average Yield
1.5 ml
1.90
2.10
13.9 µg
5.0 ml
1.90
2.10
33.0 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 5 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.
Plasmid DNA Kit Specifications
Specifications
Cat. #
PDH100
PDH300
PD100
PD300
PDL100
PDL300
96PDC04/10
96PDV04/10
FormatSpin ColumnSpin ColumnSpin Column96 Well Plate
Binding Capacity50 µg30 µg30 µg35 µg
Culture Input1-7 ml1-5 ml1-4 ml1-5 ml
Plasmid Size1-15 kb1-15 kb10-50 kb1-15 kb
Typical Yieldup to 50 µg10-30 µg10-25 µg20-35 µg/well
Elution Volume30-100 µl30-100 µl50-100 µl100 µl
Operation Time<15 minutes<15 minutes<15 minutes25 minutes
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