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DNA extraction kit for soil urine and method

A kit and soil technology, applied in the field of soil urine DNA extraction kits, can solve the problems of low DNA content and certain difficulty, and achieve the effect of overcoming adsorption and reducing DNA loss

Inactive Publication Date: 2017-08-18
ANHUI SENPENG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Urine contains a certain amount of DNA, which can be extracted by various methods and used for PCR detection, STR typing detection, and next-generation sequencing analysis; however, the DNA content in urine is low, and after the urine soaks the soil, it is The adsorbed DNA content is lower, and it is difficult to extract DNA from soil urine

Method used

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  • DNA extraction kit for soil urine and method
  • DNA extraction kit for soil urine and method
  • DNA extraction kit for soil urine and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Extraction of DNA from soil urine with silica nanoparticles.

[0035] 1) DNA extraction

[0036] Scrape the surface soil containing urine, dry it, take 100-200 mg soil sample, add it to the sample tube, then add 400ul soil lysate and 20ul proteinase K, shake and mix, heat and crack at 75°C for 15 minutes; Centrifuge at high speed for 5 minutes, transfer the supernatant to a new sample tube, add 800 ul binding solution, and mix well; transfer the mixed solution to a centrifuge tube containing 15 ul silica nanoparticles, mix well and let stand for 15 minutes ; Centrifuge at 8000rpm for 1 minute to remove the supernatant; add 500 ul of washing solution, mix well, and centrifuge at 8000rpm for 1 minute to remove the supernatant; add 500 ul of washing solution, mix well, and centrifuge at 8000rpm for 1 minute to remove the supernatant; heat at 70°C for 3 minutes; add 20 ul of eluent, shake and mix, heat at 70°C for 3 minutes; centrifuge at 8000rpm for 1 minute, an...

Embodiment 2

[0039] Example 2 Extraction of soil urine DNA with silica gel membrane

[0040] 1) DNA extraction

[0041] Scrape the surface soil containing urine, dry it, take 100-200 mg soil sample, add it to the sample tube, then add 400ul soil lysate and 20ul proteinase K, shake and mix, heat and crack at 75°C for 15 minutes; Centrifuge at high speed for 5 minutes, transfer the supernatant to a new sample tube, add 800 ul of binding solution, and mix well; transfer the mixed solution to a spin column equipped with a silica gel membrane, and centrifuge at 12,000 rpm for 1 minute; discard the waste in the collection tube solution, add 500 ul rinse solution, and centrifuge at 12,000 rpm for 1 minute; discard the waste liquid in the collection tube, add 500 ul rinse solution, and centrifuge at 12,000 rpm for 1 minute; discard the waste liquid in the collection tube, centrifuge at 12,000 rpm for 3 minutes, and transfer the spin column to In a new centrifuge tube, heat at 70°C for 3 minutes; ...

Embodiment 3

[0044] Example 3 Extraction of soil urine DNA with glass fiber membrane

[0045] 1) DNA extraction

[0046]Scrape the surface soil containing urine, dry it, take 100-200 mg soil sample, add it to the sample tube, then add 400ul soil lysate and 20ul proteinase K, shake and mix, heat and crack at 75°C for 15 minutes; Centrifuge at high speed for 5 minutes, transfer the supernatant to a new sample tube, add 800 ul of binding solution, and mix well; transfer the mixture to a spin column equipped with glass fiber, and centrifuge at 12,000 rpm for 1 minute; discard the waste in the collection tube solution, add 500 ul rinse solution, and centrifuge at 12,000 rpm for 1 minute; discard the waste liquid in the collection tube, add 500 ul rinse solution, and centrifuge at 12,000 rpm for 1 minute; discard the waste liquid in the collection tube, centrifuge at 12,000 rpm for 3 minutes, and transfer the spin column to In a new centrifuge tube, heat at 70°C for 3 minutes; add 20 ul eluent,...

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Abstract

The invention relates to a DNA extraction kit for soil urine and a method. The method comprises the following steps: 1) DNA extraction: a urine-containing soil sample is subjected to splitting decomposition with a soil lysate and proteinase K, soil and a liquid supernatant are separated centrifugally, a binding solution is added to the liquid supernatant, a silicon-based DNA adsorbing material is added after the materials are mixed, the silicon-based DNA adsorbing material and a liquid are separated, the silicon-based DNA adsorbing material is rinsed, and DNA is eluted; 2) PCR amplification and electrophoresis: DNA purified in the step 1) is subjected to PCR amplification, and DNA is detected in electrophoresis apparatus. With adoption of the kit and the method, DNA adsorption by silicon dioxide sourced from sample soil is eliminated, so that unnecessary DNA loss is reduced, and high-quality and high-yield DNA from soil urine can be obtained.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid extraction and purification, and in particular relates to a soil urine DNA extraction kit and method. Background technique [0002] Silica can reversibly bind DNA under salt and specific pH conditions, which is also the theoretical basis for the extraction of DNA by the silica bead method, magnetic bead method, and silica gel membrane method; Silica can bind to DNA through hydrogen bonds, and impurities such as proteins, lipids, and sugars can be removed because they cannot be bound. After removing chaotropic salts and increasing the pH value, the interaction between DNA and silica is weakened. , DNA is released from the surface of silica to obtain purified DNA; the soil contains a large amount of silica, therefore, when extracting DNA from soil samples, it is necessary to consider the binding of silica to DNA from the sample source, once the DNA Binding to the silica from the sample source,...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1006C12Q2523/308C12Q2527/125
Inventor 熊槐孟涛黄书琴
Owner ANHUI SENPENG BIOTECHNOLOGY CO LTD
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