Solarbio Soil Genomic DNA Extraction Kit

套件內容

筆記: Once opened, Solution A, 乙, C, D should be kept at 2-8℃. PCR Enhancer should be kept at -20℃.

產品描述

• Suitability and Efficacy:
  • Soil 基因組DNA萃取 Kit is ideal for extracting microbial DNA from extreme soil environments like cinnamon soil, mud, volcanic ash, ETC.
  • It effectively lyses bacteria and fungi, preserving microbial DNA diversity.
• Humus Removal:
  • Utilizes unique humus adsorption material to efficiently and specifically remove various humus components.
  • This process doesn’t compromise yield, and purity is significantly higher compared to the phenol-chloroform extraction method.
• DNA Yield and Integrity:
  • Extracted DNA yields are substantial, and integrity is well maintained.
  • The extracted DNA is suitable for various routine operations, including enzyme digestion, 聚合酶鍊式反應, library construction, Southern blot, ETC.

協定

Add fresh opened absolute ethanol in the Washing Buffer before use, volume is based on the label of the bottle as a reference. Put the cap back on the bottle and shake well. All centrifuge steps are performed at room temperature (15℃-25℃).

1. Method A: Weigh soil sample 0.1-0.5g, add soil into the mortar, pour the proper amount of liquid nitrogen, grind immediately, and repeat three times. When the soil turns into powder, add 450ul Solution A, and keep shaking for 1-2min until the solute is completely suspended.

Method B: Weigh soil sample 0.1-0.5g in centrifuge tube (it is better to use 2ml round bottom), add 450ul Solution A, and keep shaking for 1-2min until solute is completely suspended. 筆記: The effect of grinding with liquid nitrogen will be better.

2. Add 50ul Solution B, and invert the tube several times to mix fully (don’t shake violently). Incubate at 65℃ water bath for 6 分分鐘, 倒置, and mix each 2 分分鐘.
3. Add 100ul Solution C, and invert the tube several times to mix fully (don’t shake violently). 離心機在, 12000轉速為 10 分分鐘.
4. Transfer supernatant to a new centrifuge tube, 離心機在, 12000轉速為 10 分分鐘.
5. Add 200ul Solution D to the Adsorption Column, add supernatant centrifuged from step 4 to the Adsorption Column, mixed fully by pipette. Centrifuge at 12000rpm for 1 分分鐘.
6. Mix filtrate in Collection Tube fully, add it into Adsorption Column, and centrifuge at 12000rpm for 1 分分鐘.
7. Remove waste liquid in the Collection Tube, add 500ul Washing Buffer in the Adsorption Column, and centrifuge at 12000rpm for 10 分分鐘.
8. 重複步驟 7 twice (wash three times in total).
9. Remove waste liquid in the Collection Tube, put the Adsorption Column back into the Collection Tube, and centrifuge at 12000rpm for 2 分分鐘.
10. Dry the Adsorption Column at room temperature(15℃-25℃) for a few minutes or at 50℃ for 1 分分鐘.
11. Put the Adsorption Column in a new centrifuge tube, add 50-100ul Washing Buffer (preheat at 65℃ before use), and centrifuge at 12000rpm for 1 分分鐘.
12. Add filtrate in the centrifuge tube to the Adsorption Column, centrifuge at 12000rpm for 1 分分鐘. Liquid in the centrifuge tube is soil genomic DNA extraction solution.
13. If the PCR effect of DNA is not good, properly dilute DNA concentration or add 1/10 volume of PCR enhancer.

筆記

1. Fresh soil samples will get a higher yield. And it is better to refer to proper preservation conditions for different samples.
2. If the solution shows precipitation, redissolve at 37℃ water bath for a moment, precipitation will disappear, and it does not affect DNA extraction.
3. When taking supernatant, it should avoid taking precipitation, 否則, it will block the absorption column and affect the purity of the product.
4. Volume of the Elution buffer shouldn’t be less than 50ul, if the volume is too small, it will affect recovery It is suggested to use the Elution buffer provided with the kit, using water as an Elution buffer will lose a part of the DNA. Extracted DNA should be stored at -20℃ and avoid repeated freeze-thaw cycles to prevent DNA degradation.
5. If the product contains residual humus, it will seriously affect DNA absorbance, so it should be identified by a combination of electrophoresis detection and spectrophotometer
6. Avoid touching liquid reagents, in case of accidental contact, rinse immediately with plenty of water.

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