Presto™ Mini Plasmid Kit (PDH100, PDH300)

Mini Plasmid Kit
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. 
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
  • Spin Columns: glass fiber membrane optimized for plasmid DNA purification
  • Elution Volume: 30-100 µl
  • Storage: room temperature (15-25°C) for up to 18 months without showing any reduction in performance
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.
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
Figure 2. Sequencing data of (pBluescript) plasmid DNA purified using the Presto™ Mini Plasmid Kit.
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
Plasmid DNA Kit Specifications
Specifications  Presto™ Endotoxin Free Mini Plasmid Kit
Cat. #
PEH100
PDH100
PDH300
PD100
PD300
PDL100
PDL300
96PDC04/10
96PDV04/10
Format Spin Column Spin Column Spin Column Spin Column 96 Well Plate
Binding Capacity 40 µg 50 µg 30 µg 30 µg 35 µg
Culture Input 1-5 ml 1-7 ml 1-5 ml 1-4 ml 1-5 ml
Plasmid Size 1-15 kb 1-15 kb 1-15 kb 10-50 kb 1-15 kb
Typical Yield up to 40 µg up to 50 µg 10-30 µg 10-25 µg 20-35 µg/well
Elution Volume 30-100 µl 30-100 µl 30-100 µl 50-100 µl 100 µl
Operation Time <60 minutes <15 minutes <15 minutes <15 minutes 25 minutes
Publications
 
 
 
 

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