Simple and fast plant genome DNA extraction method

A plant genome and extraction method technology, applied in the field of plant genome DNA extraction, can solve problems such as difficult to achieve high throughput, high liquid nitrogen consumption, sample pollution, etc., to save consumption, consume less liquid nitrogen, and avoid pollution Effect

Inactive Publication Date: 2018-05-22
ZHOUKOU NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional method of relying on a mortar to grind samples requires a large amount of liquid nitrogen and a lot of manpower and time, making it difficult to achieve high-throughput and rapid results.
Due to the need for multiple sample transfers, it is easy to cause sample contamination

Method used

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  • Simple and fast plant genome DNA extraction method
  • Simple and fast plant genome DNA extraction method
  • Simple and fast plant genome DNA extraction method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Extraction of Genomic DNA from Cotton Leaves

[0041] Step 1.1: Put a 5mm steel ball in a 2mL EP tube, and place the CTAB solution in a 65°C oven to preheat;

[0042] Step 1.2: Put one end of the young cotton leaves into the EP tube containing steel balls, snap the cap to cut off the cotton tissue inside and outside the tube, and complete the sampling;

[0043] Step 1.3: Place the EP tube with cotton leaves on one end of the EP tube plate, take another EP tube plate to clamp the EP tube, put the EP tube plate with the EP tube in liquid nitrogen for 2 minutes to freeze the material thoroughly, then remove;

[0044] Step 1.4: Turn the EP tube sheet upside down to crush the cotton leaves;

[0045] Step 1.5: Add 500 microliters of preheated CTAB solution to the EP tube, turn it upside down until uniform, then place it in a 65°C oven for 20 minutes, take it out every 10 minutes and mix it upside down;

[0046] Step 1.6: Then add 500 microliters of chloroform: is...

Embodiment 2

[0052] Example 2 Extraction of Arabidopsis Leaf Genomic DNA

[0053] Step 2.1: Put two 3mm steel balls into a 2mL EP tube, and place the CTAB solution in a 65°C oven to preheat;

[0054] Step 2.2: put one end of the young leaves of Arabidopsis thaliana into the EP tube containing steel balls, snap the cap to cut off the Arabidopsis tissue inside and outside the tube, and complete the sampling;

[0055]Step 2.3: Place the EP tube with Arabidopsis leaves on one end of the EP tube plate, take another EP tube plate to clamp the EP tube, put the EP tube plate with the EP tube in liquid nitrogen for 2 minutes to freeze the material see through, and then remove;

[0056] Step 2.4: Turn the EP tubesheet upside down to crush the Arabidopsis leaves;

[0057] Step 2.5: Add 600 microliters of preheated CTAB solution to the EP tube, turn it upside down until uniform, then place it in a 65°C oven for 30 minutes, take it out every 10 minutes and mix it upside down;

[0058] Step 2.6: Then...

Embodiment 3

[0064] Example 3 Extraction of Soybean Leaf Genomic DNA

[0065] Step 3.1: Put two 3mm steel balls into a 2mL EP tube, and place the CTAB solution in a 65°C oven to preheat;

[0066] Step 3.2: Put one end of the young soybean leaves into the EP tube containing steel balls, snap the cap to cut off the soybean tissue inside and outside the tube, and complete the sampling;

[0067] Step 3.3: Place the EP tube with soybean leaves on one end of the EP tube plate, take another EP tube plate to clamp the EP tube, put the EP tube plate with the EP tube in liquid nitrogen for 2 minutes to freeze the material thoroughly, then remove;

[0068] Step 3.4: Turn the EP tube sheet upside down to crush the soybean leaves;

[0069] Step 3.5: Add 700 microliters of preheated CTAB solution to the EP tube, turn it upside down until uniform, then place it in a 65°C oven for 40 minutes, take it out every 10 minutes and mix it upside down;

[0070] Step 3.6: Then add 700 microliters of chloroform ...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a simple and fast plant genome DNA extraction method. On the basis of a CTAB extraction method, an extraction solution formula is improved; a sample grinding method is changed; the operation steps are optimized; the plant genome DNA can be fast extracted at high flux; instruments such as a grinding instrument arenot needed; the liquid nitrogen consumption is extremely low; reagents with higher danger such as phenol are not needed.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a simple and fast method for extracting plant genome DNA. Background technique [0002] With the rapid development of molecular biology research, especially the rapid development of plant molecular biology research in recent years, plant genome DNA extraction as a basic technology is used more and more frequently in plant molecular biology research. The screening of plant mutants, the identification of transgenes after plant genetic transformation, and the detection of plant genome editing all require high-throughput, fast, and convenient means of extracting plant genomic DNA. At present, high-throughput plant genome extraction mostly relies on specific instruments or kits, which are obviously impossible for some laboratories with insufficient funds. The traditional method of relying on a mortar to grind samples consumes a large amount of liquid nitrogen and requires a l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 李成伟于德水张菊廖立冰张怡徐克东刘坤谭光轩陈璨付贝贝谢家星毛洁
Owner ZHOUKOU NORMAL UNIV
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