Introduction
The GB-Clean™ Plasmid Miniprep Kit (HiPure Mini system) provides a fast, simple, and cost-effective plasmid DNA miniprep method for routine molecular biology laboratory applications. HiPure Plasmid Mini Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Plasmid DNA purified with Mini Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of tris buffer or water.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation up to 35μg plasmid DNA from 1-5ml bacterial culture |
| Applications | Enzyme digestion, sequencing, PCR, cloning, etc. |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Conventional plasmid, plasmid less than 30KB |
| Sample amount |
High copy plasmid: 1-5ml culture medium Low copy number plasmid : 5-10ml culture medium |
| Yield | 5-35µg |
| Elution volume | <span "="" style="outline: 0px; box-sizing: border-box; font-size: 16px;">≥30μl |
| Time per run | Complete 1-24 samples in 30 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 35µg |
Principle
The HiPure Plasmid procedure is based on alkaline lysis of bacterial cells followed by adsorption of DNA onto silica in the presence of high salt. The unique silica membrane used in the kit completely replaces glass or silica slurries for plasmid DNA minipreps. The procedure consists of 3 basic steps: Preparation and clearing of a bacterial lysate by alkaline method，then transfer the supernatant to column to bind DNA. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).
Advantages
- High purity - purified plasmid can be directly used in sequencing, enzyme digestion and PCR, etc.
- Fast - it takes only 1 minute to obtain supernatant by optimized solution (10 minutes for other brands)
- High yield (see comparison below)- up to 35µg plasmid can be bound in one column
Kit Contents
| Contents | P100101 | P100103 |
| Purification Times | 50 Preps | 250 Preps |
| RNase A | 2 mg | 10 mg |
| Buffer P1 | 15 ml | 80 ml |
| Buffer P2 |
15 ml |
80 ml |
| Buffer P3 |
20 ml |
100 ml |
| Buffer PW1 |
25 ml |
140 ml |
| Buffer PW2* | 10 ml | 50 ml |
| Elution Buffer | 6 ml | 30 ml |
| HiPure DNA Mini Columns II | 50 | 250 |
| 2 ml Collection Tubes | 50 | 250 |
Storage and Stability
The kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve. After addition of RNase A，Buffer P1 is stable for 6 months when stored at 2-8°C.
Experiment Data
See also the attached data figure.
