DNA Extraction > Plasmid DNA Purification

Presto™ Mini Plasmid Kit

Presto™ Mini Plasmid Kit
Presto™ Mini Plasmid Kit

This Plasmid Kit was designed for plasmid and cosmid 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 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. Typical yields are 20-35 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 are not required and the entire procedure can be completed within 15 minutes. 

Protocol download
Advantages (Cat. # PDH100, PDH300)
  • 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
  • Plasmid glass fiber spin column (red membrane optimized for plasmd DNA purification)
  • Storage: room temperature (15-25°C) for up to 1 year
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 20 μg is obtained and the A260/A280 ratio is between 1.8-2.0. The purified plasmid (1 µg) is used in EcoR I digestion, and analyzed by electrophoresis.

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 aliquats 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 used in EcoRI digestion and analyzed by electrophoresis on a 0.8% agarose gel.
Figure 1. Plasmid DNA was extracted using the Presto™ Mini Plasmid Kit. 5 µl aliquats 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
Figure 2. Sequencing data of (pBluescript) plasmid DNA purified using the Presto™ Mini Plasmid Kit
Click image to expand
TrueBlue Lysis Buffer Optional Color Indicator
Cell Lysis
Neutralization
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.
Insufficient Mixing
Correct Mixing
Insufficient Mixing
Correct Mixing
Related Plasmid DNA Purification Products
Product Name                                                                                RXNS Catalogue Number
Presto™ Mini Plasmid Kit
100, 300
PDH100/300
100, 300
PD100/300
100, 300
PDL100/300
25
PA025
25
PIF025
25
PIF01
25
PIFE25
25
PI025
25
PIE25
10, 25
PM010/25
10, 25
PME10/25
4 x 96, 10 x 96
PDA04/10

 

 

 

 

 

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