Soil and Fecal DNA Extraction Kit
Broad-spectrum application, powerfully removes humic acid.
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Instruction manual- Product Description
- Composition
- Precautions
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Product Storage:
1. All reagent components are transported and stored at room temperature (15–25°C), with a shelf life of one year.
2. When stored at low temperatures, SDL buffer 1 may precipitate. Before use, place the entire reagent bottle in a 55°C water bath until the precipitate is completely dissolved, then mix thoroughly before use.
Product Overview:
This product is suitable for extracting genomic DNA from a wide range of samples, including soil, feces, water samples, and membrane filters. By combining silica‑based column purification with an efficient humic acid removal protocol, this kit effectively eliminates inhibitory substances such as humic acids, yielding high‑quality DNA. The resulting DNA is compatible with various downstream applications, including PCR, restriction digestion, and library construction.
Pre-experiment preparation
As indicated in the table below, dilute Washing Buffer 1 and Washing Buffer 2 with anhydrous ethanol; store the diluted solutions at room temperature.
Component Amount of anhydrous ethanol added Washing Buffer 1 12 mL Washing Buffer 2 64 mL User-provided
Desktop high-speed centrifuge, vortex mixer, anhydrous ethanol, 2 mL centrifuge tubes.
Instructions for Use
1. Weigh 200 mg of glass bead 1 and 200 mg of glass bead 2 into separate 2-mL centrifuge tubes, then add the sample.
a. Soil/Fecal Samples: Add 100–500 mg of soil sample, then add 800 μL of SDL Buffer 1 and 120 μL of SDL Buffer 2.
b. Filter membrane sample: Add the shredded filter membrane, then add 800 μL of SDL buffer 1 and 120 μL of SDL buffer 2.
c. Liquid sample: Add 200 μL of the liquid sample, then add 600 μL of SDL buffer 1 and 120 μL of SDL buffer 2.
2. Vortex or homogenize the above mixture; recommended vortexing conditions: maximum speed for 10 min; homogenization conditions: 50 Hz for 5 min. If higher DNA integrity is required, milder grinding conditions may be used; recommended vortexing conditions: medium speed for 10 min; homogenization conditions: 25 Hz for 10 min; however, reduced grinding intensity may compromise nucleic acid yield.
3. Add 200 μL of SDL buffer 3 to the sample that has been vortexed or homogenized, and manually shake to mix for 10 seconds.
4. Centrifuge at room temperature at 13,000 rpm (15,000 × g) for 2 min, then transfer 700 μL of the supernatant to a new 1.5 mL centrifuge tube.
5. Add 140 μL of HAR Buffer 2B, vortex to mix thoroughly, centrifuge at 13,000 rpm (15,000 × g) for 5 min, and transfer 700 μL of the supernatant into a new 2 mL centrifuge tube.
6. Add an equal volume of DNA Binding Buffer and vortex to mix thoroughly.
7. Place the DNA adsorption column into the collection tube, transfer the mixture from step 6 into the binding column, and centrifuge at 13,000 rpm (15,000 × g) for 30 seconds; discard the supernatant.
8. Repeat step 7 until all the mixture has passed through the binding column.
9. Place the DNA adsorption column into the collection tube, add 500 μL of Washing Buffer 1 to the column, centrifuge at 13,000 rpm (15,000 × g) for 30 seconds, and discard the supernatant.
10. Place the DNA adsorption column into the collection tube, add 700 μL of Washing Buffer 2 to the column, centrifuge at 13,000 rpm (15,000 × g) for 30 seconds, and discard the supernatant.
11. Repeat step 10.
12. Place the DNA adsorption column into the collection tube and spin at 13,000 rpm (15,000 × g) for 2 minutes with no load.
13. Place the DNA adsorption column into a new 1.5 mL centrifuge tube, add 50–100 μL of Elution Buffer to the column, incubate at room temperature for 1–2 minutes, and then centrifuge at 13,000 rpm (15,000 × g) for 1 minute to elute the DNA.
14. DNA should be stored at −20°C.
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Product Composition
Item number Component Specifications 400-210A SDL buffer 1 44 mL 400-210B SDL buffer 2 7 mL 400-210C SDL buffer 3 11 mL 400-210D HAR Buffer 2B 8 mL 400-210E DNA Binding Buffer 39 mL 400-210F Washing Buffer 1 18 mL 400-210G Washing Buffer 2 16 mL 400-210H Elution Buffer 6 mL 400-210I DNA adsorption column (with collection tube) 50 sets 400-210J Glass bead 1 14 g 400-210K Glass beads 2 14 g -
Precautions
1. Before first use, please add the specified amounts of anhydrous ethanol to Washing Buffer 1 and Washing Buffer 2, respectively. After adding, promptly check the box to indicate that ethanol has been added, to avoid adding it multiple times.
2. When using this product, be sure to take appropriate personal protective measures, such as protective clothing, gloves, goggles, and face shields.
3. This product is intended for research use only; do not use for diagnostic purposes.
Keywords
GOONIE
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Goonie Biotechnology (Guangzhou) Co., Ltd.
Room 321–332, Building G5, No. 39 Ruihe Road, Huangpu District, Guangzhou
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