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Method for extracting and purifying DNA of plants like cactus

A purification method and cactus technology, applied in chemical instruments and methods, sugar derivatives, sugar derivatives, etc., can solve the problems of pure DNA, can not be obtained, and achieve low cost, overcome interference, improve yield and purity. Effect

Inactive Publication Date: 2008-05-21
GUIZHOU FRUIT INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, cacti plants contain mucus substances, and DNA extraction using the standard CTAB method will be interfered by mucus substances, and pure DNA cannot be obtained, which cannot meet the needs of research.

Method used

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  • Method for extracting and purifying DNA of plants like cactus

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1: adopt following method to extract dragon fruit DNA:

[0039] (1) Collect the tender stems of dragon fruit, rinse them with clean water, dry them in air, take 5-8g and quickly grind them into powder in liquid nitrogen, and then quickly transfer them to a 1.5ml centrifuge tube;

[0040] (2) Immediately add 600 μl of cell lysate preheated at 65°C, place it in a water bath at 65°C and incubate for 1 hour, shake gently once every 5-10 minutes, centrifuge the lysed sample for 20 minutes, and pipette the supernatant into a centrifuge tube middle;

[0041] (3) Add 4°C saturated phenol solution and normal temperature chloroform (saturated phenol solution: chloroform = 1:1) with the same volume as the supernatant to the centrifuge tube of step (2), shake gently for 10 minutes, and centrifuge at 12000r / min for 20 minute;

[0042](4) Take the supernatant into a 1.5ml centrifuge tube, add 4°C potassium acetate solution 0.1 times the volume of the supernatant and -20°...

Embodiment 2

[0046] Embodiment 2: Adopt the following method to extract cactus DNA:

[0047] (1) Collect the leaves of cactus, rinse them with clean water, dry them, take 5-8g and quickly grind them into powder in liquid nitrogen, and then quickly transfer them to a 1.5ml centrifuge tube;

[0048] (2) Immediately add 600 μl of cell lysate preheated at 65°C, place in a water bath at 65°C and incubate for 70 minutes, shake gently once every 5-10 minutes, centrifuge the lysed sample for 22 minutes, and pipette the supernatant into a centrifuge tube middle;

[0049] (3) Add 4°C saturated phenol solution and normal temperature chloroform with the same volume as the supernatant to the centrifuge tube in step (2), shake gently for 8 minutes, and centrifuge at 12000r / min for 22 minutes;

[0050] (4) Take the supernatant into a 1.5ml centrifuge tube, add 4°C potassium acetate solution 0.1 times the volume of the supernatant and -20°C isopropanol of the same volume as the supernatant, and place in ...

Embodiment 3

[0054] Embodiment 3: adopt following method to extract dragon fruit DNA

[0055] (1) Collect the tender stems of dragon fruit, rinse them with clean water, dry them in air, take 5-8g and quickly grind them into powder in liquid nitrogen, and then quickly transfer them to a 1.5ml centrifuge tube;

[0056] (2) Immediately add 600 μl of cell lysate preheated at 65°C, place in a water bath at 65°C and incubate for 50 minutes, shake gently once every 5-10 minutes, centrifuge the lysed sample for 18 minutes, and pipette the supernatant into a centrifuge tube middle;

[0057] (3) Add 4°C saturated phenol solution and normal temperature chloroform with the same volume as the supernatant to the centrifuge tube in step (2), shake gently for 12 minutes, and centrifuge at 12000r / min for 18 minutes;

[0058] (4) Take the supernatant into a 1.5ml centrifuge tube, add 4°C potassium acetate solution 0.1 times the volume of the supernatant and -20°C isopropanol of the same volume as the super...

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Abstract

The invention provides an extraction and purification method of opuntia DNA and is especially fit for extracting and purifying the DNA from the muciferous opuntia. The method of the invention is an improvement based on a standard CTAB method and adopts the methods of adjusting the test order, properly prolonging the extraction time of organic solvent, increasing the extraction times etc. to effectively conquer the interference of mucus substance during the DNA extraction process, effectively remove the interference substances in extract, such as sugar, phenol, enzyme etc. and improves the yield and the purity of the DNA extraction, and the obtained DNA extract can meet the requirement of further research. The whole extraction and purification method does not require other special instrument and has low cost and simple and convenient operation, and general testing personnel can achieve successfully.

Description

technical field [0001] The invention relates to a method for extracting and purifying DNA, in particular to a method for extracting and purifying DNA of cactus plants that produce mucus during DNA extraction. Background technique [0002] Cacti are mainly distributed in the tropical regions of America. There are more than 2,000 known species in the world, among which Mexico is the most. There are also a few wild cactus plants in the southwestern and southern coastal areas of my country. Over the past ten years, through introduction and cultivation, cactus plants have increased year by year in my country. Mainly distributed in southern my country. Due to the increasing market demand for edible and fruiting cactus plants, in order to better utilize and protect these resources, a lot of research work is being carried out, including revealing their interspecific relationship, genetic diversity, and molecular marker breeding. , the premise of these studies is to get DNA with hi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H21/04C07H1/08
Inventor 蔡永强陶刚向青云刘涛金吉芬韦茜
Owner GUIZHOU FRUIT INST
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