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Description
The Water DNA Isolation Kit extracts high-purity genomic DNA from microorganisms in river, lake, pond, aquaculture, tap and seawater samples. Microorganisms are first collected on a 47 mm, 0.22–0.45 µm filter membrane, so the kit handles sample volumes from 10 mL up to 30 L, from clear tap water to turbid pond water. It is suited to environmental DNA (eDNA) studies, water quality monitoring and qPCR detection of specific bacteria.
Water samples carry PCR inhibitors such as humic acid and metal ions that can carry over into the DNA and cause failed or delayed amplification. The kit combines lysozyme and protease lysis with a DNA-Only silica membrane spin column and optimized wash buffers to remove these inhibitors, with no phenol/chloroform extraction and no ethanol or isopropanol precipitation.
The DNA-Only column binds DNA but not RNA, so no RNase needs to be added. All centrifugation steps run at room temperature in a benchtop centrifuge, and 24 samples can be processed in about 40 minutes after filtration. Purified DNA is ready for PCR, qPCR, restriction digestion, Southern blotting and library construction.
Specifications
| Item | Specifications |
|---|---|
| Product Type | Spin-column DNA extraction kit for water samples |
| Preps | 50 |
| Sample Input | Microorganisms collected on a 47 mm, 0.22–0.45 µm filter membrane |
| Water Volume per Sample | 10 mL to 30 L, depending on turbidity and microbial content |
| Sample Types | River, lake, pond, aquaculture pond, tap water and seawater |
| Column | DNA-Only silica membrane spin column |
| Column Binding Capacity | 80 µg DNA |
| Column Loading Volume | 800 µL |
| Elution Volume | 50–200 µL |
| Processing Time | About 40 minutes for 24 samples, after filtration |
| Throughput | 1–24 samples per run |
| Centrifuge | Benchtop centrifuge at room temperature, up to ~17,000 × g |
| Typical Purity | OD260/280 1.7–1.9 |
| Downstream Applications | PCR, qPCR, restriction digestion, Southern blotting, library construction |
Typical DNA Yield and Purity
| Water Source | Volume Filtered | DNA Yield | OD260/280 | OD260/230 |
|---|---|---|---|---|
| River water | 400 mL | 1–2 µg | 1.7–1.9 | 1.7–1.9 |
| Pond water | 400 mL | 5–7 µg | 1.7–1.9 | 1.7–1.9 |
| Aquaculture pond water | 20 mL | 1–2 µg | 1.7–1.9 | 1.8–1.9 |
| Tap water | 4,000 mL | 0.05–0.1 µg | ≈1.8 | 1.9–2.0 |
Reference data. Actual yield depends on the water source, turbidity and microbial content.
Features
- Large sample range: Filter 10 mL to 30 L of water onto a single membrane, from clear tap water to turbid pond water.
- Inhibitor removal: Silica column chemistry removes humic acid, metal ions and salts for reliable PCR and qPCR.
- No RNase needed: The DNA-Only column excludes RNA, so no RNase is added and labs avoid RNase contamination.
- Fast: About 40 minutes for 24 samples after filtration.
- Room-temperature workflow: All centrifugation runs at room temperature; no 4 °C centrifuge or alcohol precipitation.
- Safer chemistry: No phenol or chloroform extraction.
- High-quality DNA: Large DNA fragments, OD260/280 1.7–1.9, low ion content.
Applications
- Environmental DNA (eDNA) extraction for biodiversity and ecological surveys.
- qPCR detection of specific bacteria in water, such as sulfate-reducing bacteria with the Biofargo Sulfate-Reducing Bacteria Probe qPCR Kit.
- Microbial community analysis and 16S rRNA gene sequencing of water samples.
- Water quality monitoring of rivers, lakes, reservoirs and tap water.
- Aquaculture pond monitoring and pathogen screening.
- Seawater and marine samples collected by membrane filtration.
Kit Components
| Component | DE-05613 (50 Preps) |
|---|---|
| Buffer SG1 | 35 mL |
| Buffer SG2 | 2 mL |
| Buffer SG3 | 35 mL |
| Buffer SG4 | 1.2 mL |
| Buffer PW | 30 mL |
| Buffer WB | 25 mL (add 60 mL absolute ethanol before first use) |
| Buffer EB | 15 mL |
| Buffer TE | 60 mL |
| Protease | 1.8 mL |
| Lysozyme | 250 mg |
| DNA-Only Spin Columns | 50 |
| Instructions for Use | 1 |
Required but Not Included
- Filtration apparatus and 47 mm filter membranes, 0.22–0.45 µm pore size
- Absolute ethanol (96–100%)
- 1.5 mL and 2 mL microcentrifuge tubes
- Shaking incubator at 37 °C (180 rpm)
- Water bath or heat block at 65 °C
- Benchtop microcentrifuge (~17,000 × g)
- Sterile scissors and forceps for cutting the membrane
Storage & Shipping
- Store the kit dry at room temperature (15–25 °C) for up to 12 months, or at 2–8 °C for longer storage.
- If buffers precipitate after cold storage, warm them at 37 °C for 10 minutes and mix before use.
- Protease: stable at room temperature for 3 months; store at 4 °C for best stability. Do not freeze.
- Lysozyme: store at –20 °C. After dissolving, divide into aliquots and store at –20 °C.
- Ships at ambient temperature.
FAQs
Can this kit extract DNA from seawater?
Yes. Filter the seawater through a 47 mm, 0.22–0.45 µm membrane according to the protocol, then process the membrane with the kit. The column wash steps remove salts and other inhibitors before elution.
How much water should I filter?
Anywhere from 10 mL to 30 L per sample, depending on turbidity and microbial content. Use smaller volumes for turbid water and larger volumes for clean water such as tap water.
Is the DNA suitable for qPCR?
Yes. Inhibitors are removed during column purification, so the DNA can be used directly for PCR and qPCR, for example with the Biofargo Sulfate-Reducing Bacteria Probe qPCR Kit.
Do I need to add RNase?
No. The DNA-Only spin column does not bind RNA, so no RNase is required.
What equipment do I need?
A filtration setup with 47 mm membranes, a 37 °C shaking incubator, a 65 °C water bath or heat block, a benchtop microcentrifuge, and absolute ethanol.
How long does the extraction take?
About 40 minutes for 24 samples after the water has been filtered.
What DNA yield can I expect?
Yield depends on the water source and microbial load. Typical results are 1–2 µg from 400 mL of river water and 5–7 µg from 400 mL of pond water.
