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Product Overview
In this kit, plasmid DNA is purified rapidly by modified SDS alkaline lysis combined with selective adsorption of DNA on a DNA preparation membrane. It applies to extract up to 20 µg of high purity plasmid DNA from 1–5 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.
Cells from 1–5 mL of culture are lysed with 250 µL each of Solutions A, B and C. The cleared lysate is treated with one tenth volume of Endo-Remove Buffer, held in an ice bath for 10 min and incubated at 42°C for 5–10 min, then mixed with three volumes of absolute ethanol and bound to Adsorption column P. The column is washed twice with 600 µL of Wash Buffer and eluted in 30–100 µL, with 50 µL recommended. Purified plasmid DNA has an OD260/OD280 of about 1.8–2.0.
Specifications
| Format | Spin column (Adsorption column P) |
|---|---|
| Lysis method | Modified SDS alkaline lysis |
| Special treatment | Endo-Remove Buffer treatment step |
| Sample type | Bacterial culture |
| Sample input | 1–5 mL bacterial culture |
| Yield | Up to 20 µg plasmid DNA |
| Processing time | About 1 h |
| Purity | OD260/OD280 is about 1.8–2.0 |
| Elution volume | 30–100 µL, 50 µL recommended |
Kit Components
| Component | 50 T | 200 T |
|---|---|---|
| Solution A | 15 mL | 60 mL |
| Solution B | 15 mL | 60 mL |
| Solution C | 15 mL | 60 mL |
| Endo-Remove Buffer | 4 mL | 16 mL |
| Wash Buffer | 60 mL | 240 mL |
| Elution Buffer | 5 mL | 20 mL |
| RNase A | 75 µL | 300 µL |
| Adsorption column P | 50 set | 200 set |
Applications
- Plasmid preparation for sequencing
- Restriction enzyme digestion and ligation
- PCR
- Transformation and transfection
- Cell microinjection and gene silencing experiments
Key Features
- Fast and high yield: Up to 20 µg plasmids can be 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
Solutions A, B and C, Wash Buffer, Elution Buffer and the P columns 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
- Invert 5–10 times after adding Solution B and keep this lysis step to 5 min or less.
- Hold the lysate in an ice bath for 10 min after adding Endo-Remove Buffer, then incubate at 42°C for 5–10 min.
- Carry out the centrifugation that follows the Endo-Remove step at 20°C or above.
- Add three volumes of absolute ethanol before loading the column.
- Open the column for 2 min at room temperature to remove residual ethanol, then elute with buffer at 56°C.
- The manufacturer does not publish an endotoxin specification for this kit; verify the endotoxin level in-house if the application requires a defined level.
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.
Connexin 43 overexpression induces lung cancer angiogenesis in vitro following phosphorylation at Ser279 in its C‑terminus. Oncology Letters, 2022. 10.3892/ol.2022.13413
NSUN4-mediated m5C RNA methylation protects retinal cells against excitotoxic injury via the SHH signaling pathway. PLoS One, 2026. 10.1371/journal.pone.0347414
