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High-Speed Plasmid Mini Kit (PD100, PD300)

DNA Extraction/Plasmid DNA Purification

The High-Speed Plasmid Mini Kit was designed for both plasmid and cosmid DNA purification. This plasmid kit will purify plasmid DNA/cosmid DNA from 1-5 ml of cultured bacterial cells using an efficient plasmid miniprep system. The High-Speed Plasmid Mini Kit uses a modified alkaline lysis method and RNase treatment to obtain clear cell lysate with minimal genomic DNA/RNA contaminants. In the presence of chaotropic salt, plasmid DNA in the lysate binds to the glass fiber matrix of the plasmid spin column. Contaminants are removed with a Wash Buffer (containing ethanol) and the purified plasmid DNA is eluted by a low salt Elution Buffer, TE or water. Typical yields are 20-30 μg for high-copy number plasmid or 3-10 μg for low-copy number plasmid from 4 ml of cultured bacterial cells. This plasmid kit does not require DNA phenol extraction or alcohol precipitation and the entire procedure can be completed within 15 minutes.

DOCUMENTS

DESCRIPTION

Specifications (Cat. # PD100, PD300)

  • Purify plasmid DNA or cosmid DNA within 15 minutes!
  • High Yield: up to 30 μg of pure plasmid DNA or cosmid DNA
  • Plasmid Size: 1-15 kb
  • Sample Volume: 1-5 ml of bacterial cells
  • Spin Columns: glass fiber membrane optimized for plasmid DNA purification
  • Elution Volume: 30-100 µl
  • Storage: dry at room temperature (15-25ºC)

APPLICATIONS

Restriction Enzyme Digestion, Library Screening, Ligation, PCR, Transformation and Sequencing Reactions

COMPONENTS

  • PD1, PD2, PD3 Buffers
  • W1 Buffer
  • Wash Buffer
  • Elution Buffer (10 mM Tris-HCl, pH8.5 at 25°C)
  • RNase A
  • PD Columns
  • 2 ml Collection Tubes

QUALITY CONTROL

The quality of the High-Speed Plasmid Mini Kit is tested on a lot-to-lot basis according to Geneaid's ISO-certified quality management system. Plasmid DNA from a 4 ml overnight E. coli (DH5α) culture, containing plasmid pBluescript (A600 > 2 U/ml) is purified using the High-Speed Plasmid Mini Kit. Following the purification process, a yield of more than 20 μg is obtained and the A260/A280 ratio is between 1.8-2.0. The purified plasmid (1 µg) is used in EcoRI digestion, and checked by electrophoresis.

https://test72.grnet.com.tw/geneaid/data/editor/images/index7.jpg

Presto™ Mini Plasmid Kit Functional Test Data

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Figure 1 .Plasmid DNA was extracted using both the High-Speed Plasmid Kit (lane 3,4) and the equivalent competitor's plasmid kit (lane 1,2). The purified supercoiled plasmid DNA [1.5 ml overnight E. coli (DH5α) culture, containing a 3 kb plasmid pBluescript (A600 > 2 U/ml)], from a 50 µl eluate was used in EcoRI digestion and analyzed by electrophoresis on a 1% agarose gel. M = Geneaid 1 Kb DNA Ladder.

 
 
Test
260/280
260/230
Yield
Competitor Q
1
1.88
2.25
8.30 µg
2
1.88
2.24
8.40 µg
Geneaid
3
1.87
2.21
8.30 µg
4
1.89
2.23
8.50 µg
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Figure 2 .Sequencing data of (pBluescript) plasmid DNA purified using the High-Speed Plasmid Mini Kit

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Figure 3 .A variety of plasmid and cosmid DNA was purified using the High-Speed Plasmid Mini Kit. Plasmid pBluescript II (lane 1), pUC18 (lane 2), pBR 322 (lane 3), pGEM with 1 kb insert (lane 4), pET 43.1 with 2.7 kb insert (lane 5) and cosmid (50 kb, lane 6) DNA from 50 µl eluates was used in EcoRI digestion and analyzed by electrophoresis on a 1% agarose gel. M1 = Geneaid 1 Kb DNA Ladder, M2 = Lambda-Hind III

Video 1 .A step by step demonstrational video of the High-Speed Plasmid Mini Kit. Plasmid DNA from a 4 ml overnight E. coli (DH5α) culture, containing plasmid pBluescript (A600 > 2 U/ml) is purified using the High-Speed Plasmid Mini Kit.

Plasmid DNA Kit Specifications

 
SpecificationsPresto™ Endotoxin Free Mini Plasmid KitPresto™ Mini Plasmid KitHigh-Speed Plasmid Mini KitLarge Plasmid DNA Extraction KitPresto™ 96 Well Plasmid Kit
Cat. #PEH100PDH100
PDH300
PD100
PD300
PDL100
PDL300
96PDC04/10
96PDV04/10
FormatSpin ColumnSpin ColumnSpin ColumnSpin Column96 Well Plate
Binding Capacity40 µg50 µg30 µg30 µg35 µg
Culture Input1-5 ml1-7 ml1-5 ml1-4 ml1-5 ml
Plasmid Size1-15 kb1-15 kb1-15 kb10-50 kb1-15 kb
Typical Yieldup to 40 µgup to 50 µg10-30 µg10-25 µg20-35 µg/well
Elution Volume30-100 µl30-100 µl30-100 µl50-100 µl100 µl
Operation Time<60 minutes<15 minutes<15 minutes<15 minutes25 minutes

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