| Field | Specification |
|---|---|
| Applications | |
| Storage | |
| Shelf life | |
| Catalog no. (Mfr.) | |
| Main SKU |
Product Overview
This kit uses a unique buffer system that combines with traditional isopropanol precipitation to quickly obtain large amounts of high-purity plasmid DNA. The plasmid DNA extracted using this kit can be applied to various routine operations, including restriction enzyme digestion, PCR, sequencing, ligation, transformation and transfection of a variety of cell experiments.
Bacterial cells are collected by centrifugation, lysed with 5 mL each of Solutions A, B and C, and the lysate is cleared through Filter column S. Plasmid DNA is precipitated with two thirds volume of isopropanol, washed twice with 6 mL of 75% ethanol and dissolved in 0.3–1 mL of Elution Buffer. From high-copy plasmids, 100 mL of culture gives about 200–1,500 µg; from low-copy plasmids, 200 mL of culture gives about 100–400 µg. Purified DNA has an OD260/OD280 of usually about 1.8–2.0.
Specifications
| Format | Filter column with isopropanol precipitation |
|---|---|
| Sample type | Bacterial culture |
| Sample input, high-copy plasmid | 100 mL bacterial culture |
| Sample input, low-copy plasmid | 200 mL bacterial culture |
| Yield, high-copy plasmid | About 200–1,500 µg |
| Yield, low-copy plasmid | About 100–400 µg |
| Purity | OD260/OD280 is usually about 1.8–2.0 |
| Elution volume | 0.3–1 mL Elution Buffer |
Kit Components
| Component | 10 T | 20 T |
|---|---|---|
| RNaseA | 250 µL | 500 µL |
| Solution A | 50 mL | 100 mL |
| Solution B | 50 mL | 100 mL |
| Solution C | 50 mL | 100 mL |
| MagPurp | 250 µL | 500 µL |
| Elution Buffer | 10 mL | 20 mL |
| Filter column S | 10 vial | 20 vial |
Applications
- Restriction enzyme digestion
- PCR
- Sequencing
- Ligation and transformation
- Transfection of cultured cells
Storage & Handling
The kit is stored at room temperature (15–25°C) for 12 months, or at 2–8°C for longer storage. RNase A is stored at -20°C for 1 year; Solutions A, B and C, MagPurp, Elution Buffer and the filter columns are stored at room temperature for 1 year. Solution A is stored at 4°C once RNase A has been added to it.
Usage Notes
- Handle Solutions B and C carefully.
- Avoid violent shaking during lysis; invert the tube gently 6–8 times.
- Add RNase A to Solution A before use and store Solution A at 4°C thereafter.
- Preheat the Elution Buffer to 65–70°C before dissolving the DNA.
- Air-dry the pellet for 10–20 min at room temperature after the ethanol washes.
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.
Itaconate inhibits ferroptosis of macrophage via Nrf2 pathways against sepsis-induced acute lung injury. Cell Death Discovery, 2022. 10.1038/s41420-021-00807-3
