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Presto™ Mini Plasmid Kit (PDH100, PDH300)

Presto™ Mini Plasmid Kit (PDH100, PDH300)

This miniprep Plasmid Kit was designed for plasmid DNA purification from 1-7 ml of cultured bacterial cells. For processing larger volumes, the Presto™ Midi Plasmid Kit is also available. Both plasmid kits include TrueBlue Lysis Buffer (an optional color indicator) to prevent common handling errors while ensuring efficient cell lysis, neutralization and maximum plasmid product yield. The Presto™ Mini Plasmid 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 mini 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. DNA phenol extraction or alcohol precipitation are not required and the entire procedure can be completed within 15 minutes.

 

DESCRIPTION

Specifications (Cat. # PD100, PD300)

  • Purify plasmid DNA within 15 minutes!
  • 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 50 µg of pure plasmid DNA
  • Sample Volume: 1-7 ml of cultured bacterial cells
  • Spin Columns: glass fiber membrane optimized for plasmid DNA purification
  • Elution Volume: 30-100 µl
  • Storage: dry at room temperature (15-25°C)

APPLICATIONS

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

COMPONENTS

  • PD1, PD2, PD3 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™ 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™ Mini Plasmid Kit. Following the purification process, a yield of more than 30 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.

 

Figure 1 .Plasmid DNA was extracted using the Presto™ Mini Plasmid Kit. 5 µl aliquots of a 100 µl eluate of purified super coiled plasmid DNA from 1.5, 3, 5 and 7 ml overnight E. coli (DH5α) culture, containing a 3 kb plasmid pBluescript and pBR322 (OD600 = 4 U/ml) were used in EcoRI digestion and analyzed by electrophoresis on a 0.8% agarose gel. M = Geneaid 1 Kb DNA Ladder pBluescript (1.=1.5 ml, 2=3 ml, 3=5 ml, 4=7 ml) pBR322 (5=1.5 ml, 6=3 ml, 7=5 ml, 8=7 ml)

 

Copy Number

Cell Culture Volume Yield (OD600 = 4.0)

1.5 ml

3 ml

5 ml

7 ml

High-Copy (pBluescript)

13-15 µg

27-29 µg

36-38 µg

40-42 µg

Low-Copy (pBR322)

4-6 µg

8-10 µg

12-14 µg

18-20 µg

 

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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