product description
This kit provides a quick and easy way to extract genomic DNA from water samples from a variety of sources. A large number of microorganisms are present in the water sample, and these microorganisms are widely used as important ecological indicators. Extracting high-purity genomic DNA from water is a very important means of detecting water environment values. The water sample contains a large number of inhibitors such as humic acid, metal ions, etc., even if the trace amount is present in the purified DNA, it will affect the downstream reaction, such as PCR, restriction enzyme digestion and the like. Therefore, the key to purifying water DNA is how to effectively remove the inhibitor in water.
The company's unique DNA-only silica gel spin column and solution formulation, combined with Foregene Protease, can effectively remove various inhibitors in water, without organic solvent extraction or ethanol and isopropanol precipitation, within 40 minutes Complete the extraction of DNA from the water sample.
Features
ïµ No RNase contamination: RNA in genomic DNA can be removed without additional RNase addition, avoiding exposure to exogenous RNase contamination in the laboratory.
速度 Fast speed: easy to operate, water genomic DNA extraction can be completed in 40 minutes.
方便 Convenient: Centrifugal operation is at room temperature, no need to centrifuge at 4 °C or ethanol to precipitate DNA.
ïµ Safety: no organic reagent extraction is required.
ïµ High quality: The extracted genomic DNA fragments are large, RNA-free, RN-free, and extremely low in ion content, which can meet the requirements of various experiments.
Steps
Please add absolute ethanol to Buffer WB before use. Please refer to the label on the bottle for the volume.
1. Use a filter with a diameter of 47 mm and a pore size of 0.22-0.45 μm to filter the water sample. The filtration volume depends on the turbidity and microbial content of the water sample. Scrub 1/2 or the entire filtered filter with scissors and place in a 2 ml centrifuge tube for subsequent lysis steps. Note: If the filter is not cut as much as possible, it will affect the yield and purity of genomic DNA.
2. Add 1 ml Buffer TE and 50 μl Lysozyme (see page 3 for the preparation method) to the centrifuge tube, and mix well, and shake at 37 ° C for 15 min (rotation speed: 180 rpm / min).
3. Centrifuge for 3 min at 13,300 rpm (~17,000 x g) and aspirate the supernatant with a pipette.
Note: The residual supernatant should be absorbed as much as possible to avoid affecting subsequent operations.
4. Add 600 μl BufferSG1 to the centrifuge tube with the precipitate, mix well upside down, add 30 μl Foregene Protease, 30 μl Buffer SG2, and mix thoroughly by inverting. Note: Please check Buffer SG2 for precipitation before use. If there is precipitation, please incubate the solution at 65 °C until the precipitate is dissolved.
5. Place the centrifuge tube in a 65 ° C water bath or metal bath for 5 min. Mix the tube vertically and upside down vigorously for 5 sec. The sample in the centrifuge tube is free of agglomeration to fully react with the lysate. Otherwise, it will affect the yield and purity of the DNA.
6. Add 600 μl Buffer SG3 to the centrifuge tube, mix well upside down, place in a 65 ° C water bath or metal bath for 5 min, and mix thoroughly upside down.
7. Centrifuge at 13,300 rpm (~17,000×g) for 3 min, transfer the supernatant to a new 2 ml centrifuge tube with a pipette, and avoid precipitation.
8. Add 20 μl Buffer SG4 to the centrifuge tube containing the supernatant, 280 μl of ethanol (96-100%), vortex and mix for 10 s, and collect the droplets attached to the tube cap and tube wall by transient centrifugation.
9. Place the spin column into the collection tube and add 800 μl of the mixture to a spin column, 12,000 rpm (~13,400 × g), and centrifuge for 1 min.
10. Pour off the waste liquid from the collection tube, place the spin column back into the collection tube, and add the remaining mixture to the spin column at 12,000 rpm (~13,400 × g) and centrifuge for 1 min.
11. Pour off the waste liquid from the collection tube, place the spin column back into the collection tube, add 500 μl Buffer PW to the spin column, 12,000 rpm (~13,400 × g), and centrifuge for 1 min.
12. Pour off the waste liquid from the collection tube, place the spin column back into the collection tube, add 700 μl Buffer WB to the spin column, 12,000 rpm (~13,400×g), and centrifuge for 1 min.
13. Repeat step 12 once. 14. Drain the waste from the collection tube and return the spin column to the collection tube and centrifuge at 12,000 rpm (~13,400 x g) for 1 min.
15. Transfer the spin column to a new 2 ml centrifuge tube and add 50-200 μl of Buffer EB pre-warmed at 65 °C to the center of the membrane, place at room temperature for 5 min, centrifuge at 12,000 rpm (~13,400 x g) for 1 min.
Note: If you want to increase the concentration of DNA, you can add the solution from the first centrifugation back to the spin column and centrifuge at 12,000 rpm (~13,400 × g) for 1 min.
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Blood Collection Set, Safety Blood Collection, Blood Collection Device, Blood Collection Needle with Holder, Blood Collection set with Holder, Butterful Needle with Holder
Luck Medical Consumables Co.,LIMITED , https://www.luckmedical.com