Large Plasmid Mini Kit (PDL100, PDL300)

Large Plasmid DNA Extraction Kit (10-50 kb plasmid DNA)
Large Plasmid Mini Kit (PDL100, PDL300)

The Large Plasmid DNA Extraction Kit was designed for plasmid and cosmid DNA purification from 1-4 ml of cultured bacterial cells. This plasmid kit is optimized for extracting 10-50 kb plasmid DNA using a miniprep spin column system. The Large Plasmid DNA Extraction Kit uses a modified alkaline lysis method and RNase treatment to obtain clear cell lysate with minimal genomic DNA and 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 or TE. 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. DNA phenol extraction or alcohol precipitation is not required and the entire procedure can be completed within 15 minutes. The purified plasmid DNA is ready for use in Restriction Enzyme Digestion, Ligation, PCR, and sequencing reactions.

 

 

Advantages (Cat. # PDL100, PDL300)  
  • Purify 10-50 kb plasmid DNA within 15 minutes!
  • High Yield: up to 30 μg of plasmid DNA or cosmid DNA
  • Sample: 1-4 ml of bacterial cells (10-50 kb plasmid)
  • Spin Columns: glass fiber membrane optimized for large plasmid DNA purification
  • Elution Volume: 50-100 µl
  • Storage: dry at room temperature (15-25ºC)
Applications
 
Restriction Enzyme Digestion, Library Screening, Ligation, PCR, Transformation and Sequencing Reactions
 
Components
  • PDL1, PDL2, PDL3 Buffers
  • WL1 Buffer
  • Wash Buffer
  • Elution Buffer (10 mM Tris-HCl, pH8.5 at 25°C)
  • RNase A
  • LP Columns
  • 2 ml Collection Tubes
Quality Control
 
The quality of the Large Plasmid DNA Extraction Kit is tested on a lot-to-lot basis according to Geneaid's ISO-certified quality management system. 5 µl aliquots of a 100 µl eluate of purified super coiled plasmid DNA from a 4 ml overnight E. coli (DH5α) culture, containing a 17.4 kb low copy plasmid (OD600 = 3.5 U/ml) are analyzed by electrophoresis on a 0.8% agarose gel
 
Large Plasmid DNA Extraction Kit Functional Test Data

Figure 1. Plasmid DNA was extracted using the Large Plasmid DNA Extraction Kit. 5 µl aliquots of a 100 µl eluate of purified super coiled plasmid DNA from a 4 ml overnight E. coli (DH5α) culture, containing a 17.4 kb low copy plasmid pBluescript (OD600 = 3.5 U/ml) were used in EcoRI digestion and analyzed by electrophoresis on a 0.8% agarose gel. M = Geneaid 1 Kb DNA Ladder
Test Conc. 260/280 260/230 Yield
1 87 ng/µl 1.90 2.16 8.7 µg
2 79 ng/µl 1.90 2.14 7.9 µg
3 86.4 ng/µl 1.89 2.17 8.6 µg
Plasmid DNA Kit Specifications
Specifications  Presto™ Endotoxin Free Mini Plasmid Kit
Large Plasmid DNA Extraction KIt
Cat. #
PEH100
PDH100
PDH300
PD100
PD300
PDL100
PDL300
96PDC04/10
96PDV04/10
Format Spin Column Spin Column Spin Column Spin Column 96 Well Plate
Binding Capacity 40 µg 50 µg 30 µg 30 µg 35 µg
Culture Input 1-5 ml 1-7 ml 1-5 ml 1-4 ml 1-5 ml
Plasmid Size 1-15 kb 1-15 kb 1-15 kb 10-50 kb 1-15 kb
Typical Yield up to 40 µg up to 50 µg 10-30 µg 10-25 µg 20-35 µg/well
Elution Volume 30-100 µl 30-100 µl 30-100 µl 50-100 µl 100 µl
Operation Time <60 minutes <15 minutes <15 minutes <15 minutes 25 minutes
Related Plasmid DNA Purification Products
Product Name RXNS Catalogue Number
Presto™ Endotoxin Free Mini Plasmid Kit 100 PEH100
100, 300
PDH100/300
100, 300
PD100/300
Large Plasmid DNA Extraction KIt
100, 300
PDL100/300
25
PA025
25
PIF025
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