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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.
100,00 € – 250,00 € excl. VAT
Restriction Enzyme Digestion, Library Screening, Ligation, PCR, Transformation and Sequencing Reactions
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.
| 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 |
Figure 2 .Sequencing data of (pBluescript) plasmid DNA purified using the Presto™ Mini Plasmid Kit.
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.
| Specifications | Presto™ Endotoxin Free Mini Plasmid Kit | Presto™ Mini Plasmid Kit | High-Speed Plasmid Mini Kit | Large Plasmid DNA Extraction Kit | Presto™ 96 Well 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 | 25-35 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 |
