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Presto™ Mini Plasmid Kit (PDH100, PDH300)

DNA Extraction/Plasmid DNA Purification

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.

DOCUMENTS

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.

Presto™ Mini Plasmid Kit Interactive Protocol

Presto™ Mini Plasmid Kit Functional Test Data

E. coli Strain
Cell Culture Volume Yield (OD600 = 4.0)
1.5 ml
3 ml
5 ml
7 ml
DH5α
13-15 µg
27-29 µg
36-38 µg
40-42 µg
Top 10
10-12 µg
18-20 µg
25-27 µg
27-29 µg
BL21 (DE3)
7-9 µg
13-15 µg
17-19 µg
20-22 µg

Table 1 .Plasmid DNA was extracted using the Presto™ Mini Plasmid Kit. 10 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, TOP10 and BL21(DE3) cultures, containing a 3 kb plasmid pBluescript (OD600 = 4 U/ml) were analyzed by spectrophotometer.

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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

Presto™ Mini Plasmid Kit Sequencing Data

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Figure 2 .Sequencing data of (pBluescript) plasmid DNA purified using the Presto™ Mini Plasmid Kit.

TrueBlue Lysis Buffer Optional Color Indicator

Cell Lysis
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    Correct Mixing
  • *
    Insufficient Mixing
Neutralization
  • *
    Correct Mixing
  • *
    Insufficient Mixing
Figure 3 .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

RELATED PRODUCTS

 
Product NameRXNSCatalogue Number
Presto™ Endotoxin Free Mini Plasmid Kit100PEH100
Presto™ Mini Plasmid Kit100, 300PDH100/300  
High-Speed Plasmid Mini Kit100, 300PD100/300
Large Plasmid DNA Extraction Kit100, 300PDL100/300
High-Speed Plasmid Advance Kit (50-100 ml)25PA025
Presto™ Midi Plasmid Kit25PIF025
Presto™ Midi Plasmid Kit (Endotoxin Free)25PIFE25
Geneaid™ Midi Plasmid Kit25PI025
Geneaid™ Midi Plasmid Kit (Endotoxin Free)25PIE25
Presto™ Plasmid DNA Concentration Kit25, 50, 100PC0250/500/1000
Geneaid™ Maxi Plasmid Kit10, 25PM010/25
Geneaid™ Maxi Plasmid Kit (Endotoxin Free)10, 25PME10/25
Presto™ 96 Well Plasmid Kit4 x 96, 10 x 9696PDC04/10
96PDV04/10
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