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Product Overview
This kit uses a high-capacity DNA preparation membrane that selectively adsorbs plasmid DNA, allowing rapid purification. It applies to extract up to 60 µg of high purity plasmid DNA from 5–30 mL bacterial culture. It has been used in molecular biology experiments such as sequencing, in vitro transcription and translation, restriction endonuclease digestion and bacterial transformation.
Bacteria cultured for 12–16 h are lysed with 500 µL of Solution A, 500 µL of Solution B and 700 µL of Solution C2, and the lysate is cleared by centrifugation at 12,000 rpm for 10 min. The supernatant is bound to Adsorption column P, washed twice with 600 µL of Wash Buffer and eluted in 100–200 µL of Elution Buffer. Purified plasmid DNA has an OD260/OD280 of about 1.8–2.0.
Specifications
| Format | Spin column (Adsorption column P) |
|---|---|
| Sample type | Bacterial culture |
| Sample input | 5–30 mL bacterial culture |
| Yield | Up to 60 µg plasmid DNA |
| Recovery | More than 85% of plasmid DNA from bacteria |
| Purity | OD260/OD280 is about 1.8–2.0 |
| Elution volume | 100–200 µL (at least 100 µL) |
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 |
| Elution Buffer | 10 mL | 40 mL |
| RNase A | 125 µL | 500 µL |
| Adsorption column P | 50 set | 200 set |
Applications
- Plasmid preparation for sequencing
- In vitro transcription and translation
- Restriction endonuclease digestion
- Bacterial transformation
Key Features
- Large amount: 5–30 mL of bacterial solution can be processed at one time, which improves the efficiency of the experiment.
- Fast: Few steps, simple operation, save time.
- High efficiency: More than 85% of plasmid DNA can be extracted from bacteria.
Storage & Handling
Most components are stored at room temperature for 1 year. RNase A is stored at -20°C for 1 year, and Solution A is stored at 4°C after RNase A has been added to it.
Usage Notes
- Add RNase A to Solution A before first use and store Solution A at 4°C thereafter.
- Invert 5–10 times after adding Solution B and keep this lysis step to 5 min or less.
- Ensure lysis is complete before adding Solution C2; insufficient lysis lowers yield.
- Centrifuge the column for 2 min and air-dry it for 2 min to remove residual ethanol before elution.
- Elution is most efficient at pH 7.0–8.5.
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.
