product description
This kit provides a quick and easy way to extract genomic DNA from soils of various sources. There are a large number of inhibitors in soil samples such as humic acid, metal ions, etc., even if trace amounts are present in the purified DNA, they will have a downstream reaction, such as PCR, restriction enzyme digestion and the like. Therefore, the key to purifying soil DNA is how to effectively remove the inhibitory factors in the soil.
The company's unique DNA-only silica gel spin column and solution formulation, combined with Foregene Protease, can effectively remove various inhibitors in the soil without the need for organic solvent extraction or ethanol and isopropanol precipitation, within 90 minutes. Complete the extraction of DNA from soil samples.
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: easy to operate, soil genomic DNA extraction can be completed in 90 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. Weigh 0.5g of soil sample and place in a 2ml centrifuge tube. Note: If the sample is a suspension, it can be centrifuged at 13,300 rpm (17,000 × g) for 1 min, and the precipitate is collected and then weighed 0.5 g into a 2 ml centrifuge tube.
2. Add 1 ml Buffer TE and 100 μl Lysozyme (see the fifth page for preparation) to the centrifuge tube and mix well. Warm the bath at 37 ° C for 30 min (rotation speed: 180 rpm/min).
3. Centrifuge at 7,200 rpm (~5,000 x g) for 5 min (how much effect the centrifugal speed has), and pipette the supernatant. 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 50 μl Foregene Protease, 50 μl Buffer SG2, and mix thoroughly by vortexing. 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 tube in a 65 ° C water bath or metal bath for 30 min, and mix it upside down every 10 min.
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 10 min, and mix thoroughly every 4 min.
7. Centrifuge at 12,000 rpm (~13,400 xg) for 5 min. Transfer the supernatant to a new 2 ml centrifuge tube with a pipette. Avoid sedimentation.
8. Add 20 μl Buffer SG4 to the centrifuge tube, vortex and mix well with 280 μl ethanol (96-100%) for 10 s. The droplets attached to the tube cap and tube wall can be collected by instantaneous 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 100 μl of Buffer EB pre-warmed at 65 ° C to the center of the membrane, place at room temperature for 5 min, and centrifuge at 12,000 rpm (~13,400 × g) for 1 min. Suspended again empty dropwise 100μl of Buffer EB preheated to the central film, 12,000rpm (~ 13,400 × g) centrifuged 1min. The two collected eluates were combined.
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.
Emergency Blanket/sleeping Bag/emergency Tents
Dongguan City Risen Medical Products Co., Ltd. , https://www.risenppe.com