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Endotoxin Free Mini Kit (PEH100)

DNA Extraction/Plasmid DNA Purification

The Presto™ Endotoxin Free Mini Plasmid Kit was designed for transfection grade plasmid DNA purification from 1-5 ml of cultured E. coli cells using silica-membrane spin columns. A modified alkaline lysis method and RNase treatment are used to obtain clear cell lysate with minimal genomic DNA and RNA contaminants. In addition, an efficient endotoxin removal step is integrated in the procedure and endotoxins are precipitated in a proprietary reagent. TrueBlue Lysis Buffer (an optional color indicator) is included with the kit in order to prevent common handling errors, ensuring efficient cell lysis and neutralization. DNA phenol extraction or alcohol precipitation is not required and the entire procedure can be completed within 50 minutes. The purified endotoxin free plasmid DNA is ready for use in transfection, restriction enzyme digestion, ligation, PCR, and sequencing reactions.

DOCUMENTS

DESCRIPTION

Specifications (Cat. # PEH100)

  • Endotoxin Free, Transfection Grade Plasmid DNA: <0.1 EU/µg of plasmid DNA
  • Convenient: TrueBlue Lysis Buffer is an optional color indicator included with the kit to reduce common handling errors, while ensuring efficient cell lysis, neutralization and maximum plasmid product yield so you can feel confident with each and every plasmid prep.
  • Plasmid Size: 1-15 kb
  • High Yield: up to 40 µg of transfection grade plasmid DNA
  • Sample Volume: 1-5 ml of cultured bacterial cells
  • Elution Volume: 30-100 µl
  • Spin Columns: glass fiber membrane optimized for plasmid DNA purification
  • Storage: dry at room temperature (15-25°C)

APPLICATIONS

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

COMPONENTS

  • PE1, PE2, PE3, PE4, PE5 Buffers
  • TrueBlue Lysis Buffer
  • W1 Buffer
  • Wash Buffer
  • Elution Buffer (10 mM Tris-HCl, pH8.5 at 25°C)
  • RNase A
  • PDH Columns
  • 2 ml Collection Tubes

QUALITY CONTROL

The quality of the Presto™ Endotoxin Free Mini Plasmid 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 Presto™ Endotoxin Free Mini Plasmid Kit. Following the purification process, a yield of more than 25 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 analyzed by electrophoresis.

Presto™ Mini Plasmid Kit Functional Test Data

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Figure 1 .Plasmid DNA was extracted using the Presto™ Endotoxin Free Mini Plasmid Kit from a 4 ml of overnight cultured E. coli (DH5α) culture, containing pBluescript. 2 μl aliquots of a 50 μl eluate of purified super coiled plasmid were analyzed by spectrophotometer and 0.8% agarose gel.

M = Geneaid 1 Kb DNA Ladder

 
Sample
ng/µl
260/280
260/230
Yield (ng)
1
569.4
1.89
2.32
28.5
2
578.4
1.89
2.30
28.9
3
594.4
1.89
2.31
29.7

TrueBlue Lysis Buffer Optional Color Indicator

Cell Lysis
  • *
    Insufficient Mixing
  • *
    Correct Mixing
Neutralization
  • *
    Insufficient Mixing
  • *
    Correct Mixing
Figure 2 .TrueBlue Lysis Buffer is an optional color indicator included with this Plasmid Kit to eliminate common handling errors and ensure efficient cell lysis and neutralization each and every time.

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 µg25-35 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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