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Product Overview
This kit adopts an improved SDS alkaline lysis method, combined with a high-capacity membrane that selectively adsorbs DNA, for rapid plasmid purification. A dedicated Endo-Remove Buffer step removes endotoxin during purification. It is suitable for extracting up to 60 µg of high-purity plasmid DNA from 5–30 mL of bacterial culture for molecular biology experiments such as sequencing, transfection, in vitro transcription and translation, restriction endonuclease digestion, and bacterial transformation.
Bacteria are cultured at 37°C for 12–16 h and harvested by centrifugation, then lysed with Solutions A, B and C2. Endotoxin is removed with Endo-Remove Buffer using a 10 min ice bath followed by 5–10 min at 42°C. After ethanol addition the lysate is bound to the adsorption column, washed and eluted with 100–200 µL of Elution Buffer at 56°C for 5 min. For plasmid DNA obtained with the kit the OD260/OD280 ratio is about 1.8–2.0.
Specifications
| Format | Spin column (adsorption column P) |
|---|---|
| Lysis method | Improved SDS alkaline lysis |
| Sample input | 5–30 mL bacterial culture |
| Yield | 5–60 µg plasmid DNA per preparation |
| Endotoxin removal | Endo-Remove Buffer treatment step |
| RNA removal | RNase A supplied |
| Purity (OD260/OD280) | About 1.8–2.0 |
| Elution volume | 100–200 µL (at least 100 µL) |
| Processing time | About 1 h |
Kit Components
| Component | 50 T | 200 T |
|---|---|---|
| Solution A | 25 mL | 100 mL |
| Solution B | 25 mL | 100 mL |
| Solution C2 | 35 mL | 140 mL |
| Wash Buffer | 60 mL | 240 mL |
| Endo-Remove Buffer | 9 mL | 36 mL |
| Elution Buffer | 10 mL | 40 mL |
| RNase A | 125 µL | 500 µL |
| Adsorption column (P column) | 50 sets | 200 sets |
Applications
- Plasmid preparation with endotoxin removal from bacterial culture
- Plasmid DNA for sequencing and restriction endonuclease digestion
- Plasmid DNA for transfection and cell microinjection
- In vitro transcription and translation
- Bacterial transformation, ligation and gene silencing experiments
Key Features
- Fast and high yield: 5–60 µg plasmids are extracted in about 1 h.
- Wide range of applications: Suitable for enzyme digestion, PCR, sequencing, ligation, transformation, cell microinjection, gene silencing and transfection.
Storage & Handling
Solution A is stored at 4°C for 1 year. Solutions B and C2, Wash Buffer and Elution Buffer are stored at room temperature for 1 year. Endo-Remove Buffer is stored at 2–8°C for 1 year and RNase A at -20°C for 1 year.
Usage Notes
- Mix gently rather than vigorously during lysis and ensure the bacteria are completely lysed before proceeding.
- Monitor the pH of Solution C2; it should be no higher than 4.8.
- Carry out the centrifugation that follows the Endo-Remove Buffer treatment at 20°C or above.
- Add absolute ethanol to the Wash Buffer before use.
- Use at least 100 µL of Elution Buffer and elute at 56°C for 5 min.
- The manufacturer does not publish a residual endotoxin figure for this kit; verify the level in-house if the application requires a defined endotoxin limit.
Safety & Regulatory
For research use only. Not intended for clinical, diagnostic or therapeutic use. Follow the manufacturer's manual and standard laboratory safety practice.
Customization & Add-ons: Need this molecular biology reagent tailored to your workflow? We can help with bulk or custom pack sizes, alternative formats (spin column, magnetic bead or reagent-only kits; premixed or stand-alone reference dye), matched extraction, cDNA synthesis and qPCR workflows, and sourcing of related enzymes, ladders and consumables not in our catalog. Click Talk to a Scientist to submit a request, email us at support@biohippo.com, or explore our Research Services — our team will follow up with feasibility details and next steps.
