Plant leaf DNA extraction lysis buffer solution, plant leaf DNA rapid extraction method and application

A technology for lysing buffer and plant leaves, applied in the direction of DNA preparation, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems of complicated operation, many steps, cost, etc., and achieve stable PCR amplification effect and simple extraction steps , easy-to-operate effect

Pending Publication Date: 2021-06-04
阜阳华大生命科学研究院
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the conventional CTAB plant leaf DNA extraction method has many steps and complicated operations. It needs to go through steps such as freeze-drying, grinding, cracking, extraction, purification, and dissolution. Some of the steps need to use a variety of toxic organic solvents. High, complex operation, and more time-consuming

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  • Plant leaf DNA extraction lysis buffer solution, plant leaf DNA rapid extraction method and application
  • Plant leaf DNA extraction lysis buffer solution, plant leaf DNA rapid extraction method and application
  • Plant leaf DNA extraction lysis buffer solution, plant leaf DNA rapid extraction method and application

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Embodiment 1

[0035] Embodiment 1 Plant leaf DNA rapid extraction method

[0036] 1.1 The plant leaf DNA rapid extraction method of the present invention comprises the following steps:

[0037] (1) Get the fresh leaves of plants (circular leaves with a diameter of 1 cm) with a 0.6 cm hole puncher;

[0038] (2) Add 1 steel ball with a diameter of 4mm and 150 μL of lysate to each EP tube, extract the lysis buffer including 65mM Tris-HCl, 30mM EDTA, pH8.0; (the lysate needs to soak the leaves );

[0039] (3) Grind the EP tube on the tissue grinder for 20-60s and take it out;

[0040] (4) Heat the EP tube in a water bath at 85°C for 10 minutes, then take it out, and centrifuge at 10,000 rpm for 1 minute;

[0041] (5) Take the supernatant, and get it.

[0042] 1.2 The plant leaf DNA rapid extraction method of the present invention comprises the following steps:

[0043] (1) Get the fresh leaves of plants (circular leaves with a diameter of 1 cm) with a 0.6 cm hole puncher;

[0044] (2) Add...

Embodiment 2

[0054] Embodiment 2 investigates the effect of the inventive method extracting DNA

[0055] 1.1 Preparation before experiment

[0056] (1) Prepare extraction and lysis buffer: 70mM Tris-HCl, 35mM EDTA, adjust the pH to 8.0. Mix well and place at room temperature.

[0057] (2) Prepare the EP tube.

[0058] 1.2 DNA extraction from leaves

[0059] Select the fresh leaves of 6 crops of rice, millet, tomato, corn, soybean, and tobacco, and use a puncher to punch three circular holes with a diameter of 1 cm; put the selected leaves into the corresponding numbered EP tubes , a control group (CK) is set, and the samples of the CK group are all extracted according to the conventional CTAB extraction method; the experimental groups 1-3 are all extracted DNA by the method of the present invention.

[0060] The steps of conventional CTAB extraction method are:

[0061] ① After the plant leaves are ground into powder with liquid nitrogen, transfer the powder to a centrifuge tube and a...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a plant leaf DNA extraction lysis buffer solution, a plant leaf DNA rapid extraction method and application. The plant leaf DNA extraction lysis buffer solution is prepared from the following components: 65-75 mM of Tris-HCl and 30-40 mM of EDTA, and the pH of the extraction lysis buffer solution is 8.0. According to the plant leaf DNA extraction lysis buffer solution, a plant lysate is optimized, the fresh plant leaves are directly lysed, the whole extraction process only comprises one-time liquid adding, one-time heating and one-time centrifugation and is completed in one tube, other steps are omitted, after centrifugation, supernate is taken, conventional molecular biology experiment operation can be carried out, and the PCR amplification effect of extracted DNA is stable.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a plant leaf DNA extraction lysis buffer, a method for rapidly extracting plant leaf DNA and an application thereof. Background technique [0002] In plant molecular experimental work, it is often necessary to extract DNA from a large amount of material. There are mature traditional methods for plant DNA extraction, such as the CTAB method. The CTAB method is a quick and easy method for extracting plant genomic DNA. Its principle is: mechanically disrupt plant cells, then add CTAB separation buffer to dissolve the DNA, and then extract the protein with chloroform-isoamyl alcohol, and finally obtain DNA. However, the conventional CTAB plant leaf DNA extraction method has many steps and complicated operations. It needs to go through steps such as freeze-drying, grinding, cracking, extraction, purification, and dissolution. Some of the steps need to use a variety of toxic ...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003C12Q2527/125
Inventor 陈庆春黄刚陈永佳黄苹刘家劲汪杰侯军亮余慷
Owner 阜阳华大生命科学研究院
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