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Method for quickly extracting DNA from plant leaves

A technology of plant leaves and leaves, which is applied in the field of agricultural biotechnology and molecular biology, can solve the problems of many toxic chemical reagents, complex sample processing, and high reagent prices, and achieve low cost, simple sample processing, and good experimental results.

Inactive Publication Date: 2015-04-29
HARBIN INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to solve the problems of complex sample processing, many steps, long time, many components of chemical reagents that are toxic to the environment and human body, and high reagent prices in the existing plant DNA extraction methods, and provide a method for quickly extracting sugar beet total by using a small amount of leaf tissue. DNA method

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  • Method for quickly extracting DNA from plant leaves

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specific Embodiment approach 1

[0019] Specific embodiment one: a kind of method for extracting DNA rapidly from plant leaf of the present embodiment, it is carried out according to the following steps:

[0020] 1. Take the plant leaves and place them in the centrifuge tube, and grind the leaves directly in the centrifuge tube;

[0021] 2. Add 5 to 8 times the volume of the leaf extracting buffer to the centrifuge tube after grinding the leaves in step 1 to obtain a leaf tissue mixture; treat it for 15 to 30 minutes at a water bath temperature of 55°C to 65°C, and then Add a separation liquid equal to the volume of the blade mixture, perform high-speed centrifugation, take the supernatant and add an equal volume of separation liquid, and then perform high-speed centrifugation; take the supernatant and add 2 / 3 volume of sedimentation liquid, and directly perform low-speed centrifugation to remove The supernatant collects the precipitate;

[0022] 3. The precipitate collected in step 2 is added to sterilized ...

specific Embodiment approach 2

[0027] Embodiment 2: This embodiment differs from Embodiment 1 in that the plant leaves described in Step 1 are derived from leaf tissue in growth stages and organ parts. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0028] Embodiment 3: This embodiment differs from Embodiment 1 in that the leaf tissues in the growth stage and organ parts are the leaves in the vegetative growth stage, the leaves on the bolting flower branches or the top leaves of the massive roots in the reproductive growth stage. Others are the same as in the first embodiment.

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Abstract

The invention discloses a method for quickly extracting DNA from plant leaves, and relates to a method for quickly extracting beet total DNA by using leaf tissues. The method comprises the following steps: directly grinding leaves in a grinding pestle centrifuge tube, performing high-temperature water bath treatment by using an extracting buffer solution, and adding a separation medium to perform high-speed centrifugation. The method disclosed by the invention is simple in step, less in required tissue sample, less in toxic chemical reagent, low in cost, short in operation time and good in experimental effect. The obtained DNA can be directly used for PCR (polymerase chain reaction) amplification of nucleoplasm gene fragments, and applied to molecular biology tests of molecular markers, fingerprint spectrums and the like, so that a DNA nucleic acid base material is provided for agricultural biotechnology molecular breeding and researches in the field of molecular biology.

Description

technical field [0001] The invention relates to the field of agricultural biotechnology and molecular biology, and relates to a method for rapidly extracting sugar beet total DNA by using leaf tissue. Background technique [0002] Beet cultivars include foliage sugar beet, fodder beet, table beet and sugar beet, and are widely used in agricultural production. [0003] Sugar beet is one of the important sugar crops. Leaf beet and edible beet have high edible and medical value. Feed beet is one of the excellent animal feed. The application of bioengineering and biotechnology to the molecular genetic breeding and variety improvement of sugar beet germplasm resources is inseparable from the extraction of sugar beet DNA. [0004] At present, the commonly used methods for DNA extraction of plants including Beta genus are derived from the classic SDS method, CTAB method, magnetic bead method and other methods that have been improved or modified, but these methods have many steps, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 刘巧红程大友徐德昌罗成飞于歆代翠红使淑芝鲁兆新徐伟丽
Owner HARBIN INST OF TECH
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