High-Speed Plasmid Mini Kit

Miniprep Plasmid Kit
High-Speed Plasmid Mini Kit

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.

 

Protocol download

 

 

 

 

 

 

Advantages (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 
  • Sample Volume: 1-5 ml of bacterial cells
  • Spin Columns: glass fiber membrane optimized for plasmd DNA purification
  • Storage: dry at room temperature (15-25ºC) for up to 1 year

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 Plasmd 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 EcoR I digestion, and checked by electrophoresis.

High-Speed Plasmid Mini Kit Functional Test Data
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
Sequencing Data
Figure 2. Sequencing data of (pBluescript) plasmid DNA purified using the High-Speed Plasmid Mini Kit
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
High-Speed Plasmid Mini Kit Interactive Protocol Procedure
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
Specifications Geneaid™ Mini Plasmid Kit
Cat. #
PDH100
PDH300
PD100
PD300
PDL100
PDL300
96PDC04/10
96PDV04/10
PAE040/100
Format Spin Column Spin Column Spin Column 96 Well Plate Anion Exchange
Binding Capacity 50 µg 30 µg 30 µg 35 µg 25 µg
Culture Input 1-7 ml 1-5 ml 1-4 ml 1-5 ml 1-10 ml
Plasmid Size 1-15 kb 1-15 kb 10-50 kb 1-15 kb 1-15 kb
Typical Yield up to 50 µg 10-30 µg 10-25 µg 20-35 µg/well 10-25 µg
Elution Volume 30-100 µl 30-100 µl 50-100 µl 100 µl 50-100 µl
Operation Time <15 minutes <15 minutes <15 minutes 25 minutes 80 minutes
Related Plasmid DNA Purification Products
Product Name RXNS Catalogue Number
Geneaid™ Mini Plasmid Kit 40, 100 PAE40/100
100, 300
PDH100/300
100, 300
PD100/300
100, 300
PDL100/300
25
PA025
25
PIF025
25
PIF01
25
PIFE25
25
PI025
25
PIE25
Presto™ Plasmid DNA Concentration Kit 25, 50, 100 PC0250/500/1000
10, 25
PM010/25
10, 25
PME10/25
4 x 96, 10 x 96
96PDC04/10
96PDV04/10