| Field | Specification |
|---|---|
| Applications | |
| Storage | |
| Shelf life | |
| Catalog no. (Mfr.) | |
| Main SKU |
Product Overview
This product is suitable for the separation and purification of total DNA from 1–5 mL bacterial culture. After the bacteria are broken by lysozyme, they are dissolved by Buffer BL-1, and then precipitated by Buffer BL-2 to remove protein and cell debris. The genomic DNA in the centrifugal supernatant can be bound to the purification column. After washing with Buffer RP and Buffer WB to remove the proteins and PCR inhibitors remaining on the membrane, the genomic DNA is eluted with Buffer TE and can be used in various molecular biology experiments immediately.
The bacterial pellet is resuspended in 200 µL of Buffer TE, digested with 100 µL of lysozyme solution at 37°C for 30–60 min, then treated with 225 µL of Buffer BL-1 and 225 µL of Buffer BL-2 with 30 s of vortexing each. After clarification at 13,000 rpm for 2 min the supernatant is bound to the purification column, washed with 500 µL of Buffer RP and 600 µL of Buffer WB, dried at 14,000 rpm for 1 min and eluted with 50–100 µL of warm Buffer TE. The OD260/OD280 ratio should be 1.7–1.9.
Specifications
| Format | Spin column (nucleic acid purification column) |
|---|---|
| Sample type | Bacterial culture |
| Sample input | 1–5 mL bacterial culture |
| Purity (OD260/OD280) | 1.7–1.9 |
| Elution volume | 50–100 µL Buffer TE at 37°C |
| Processing time | Within 1 hour for Gram-negative bacteria |
| Special treatment | Lysozyme supplied; no proteinase K or RNase digestion step required |
Kit Components
| Component | 50 T | 200 T |
|---|---|---|
| Buffer BL-1 | 15 mL | 60 mL |
| Buffer BL-2 | 15 mL | 60 mL |
| Buffer RP | 30 mL | 120 mL |
| Lysozyme | 600 mg | 2,400 mg |
| Buffer WB | 40 mL | 160 mL |
| Buffer TE | 15 mL | 60 mL |
| Nucleic acid purification column | 50 set | 200 set |
Applications
- Total DNA extraction from bacterial culture
- Genomic DNA for PCR
- Genomic DNA for enzyme digestion
- Genomic DNA for hybridization
Key Features
- Simple and fast: Gram-negative bacteria can obtain high-quality genomic DNA within 1 hour.
- No proteinase K or RNase digestion step is required.
- Good quality: DNA can be directly used in molecular biology experiments such as PCR, enzyme digestion, and hybridization.
Storage & Handling
Buffers BL-1, BL-2, RP, WB and TE and the purification columns are stored at room temperature for 1 year. Lysozyme is stored at -20°C for 1 year.
Usage Notes
- Avoid repeated freeze-thaw cycles.
- Confirm that absolute ethanol has been added to the wash buffers before first use.
- Thick-walled bacterial species require a longer lysozyme incubation.
- RNA can be removed by an optional RNase A treatment at 1:200.
- Ensure complete lysis to avoid blocking the column.
- Store the eluted DNA at -20°C.
- Lysozyme is supplied as 600 mg (50 T) or 2,400 mg (200 T); the protocol uses 100 µL of lysozyme solution per preparation. Follow the manufacturer's manual for its preparation.
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.
Single-Chamber Microbial Fuel Cell with an Innovative Sensing Component for Real-Time Continual Monitoring of a Wide Range of Cr(VI) Concentrations in Wastewater. Biosensors, 2025. 10.3390/bios15030158
