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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.

Quick Protocol

 

 

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
TestConc.260/280260/230Yield
187 ng/µl1.902.168.7 µg
279 ng/µl1.902.147.9 µg
386.4 ng/µl1.892.178.6 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
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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