Improved method for extracting total DNA (deoxyribonucleic acid) from polysaccharide and polyphenol plant Rhododendron lapponicum

An extraction method, Rhododendron technology, applied in the field of plant life science, can solve the problems of poor DNA integrity, many impurities, and low purity, and achieve the effect of improving DNA yield, high purity, and good repeatability

Inactive Publication Date: 2016-06-29
FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI
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Problems solved by technology

[0003] In order to solve the technical problems such as more impurities, lower purity and poor DNA integrity in the DNA extraction of Rhododendron alpine by ...

Method used

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  • Improved method for extracting total DNA (deoxyribonucleic acid) from polysaccharide and polyphenol plant Rhododendron lapponicum
  • Improved method for extracting total DNA (deoxyribonucleic acid) from polysaccharide and polyphenol plant Rhododendron lapponicum

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Embodiment 1

[0030] Rhododendron lantana (sample 1), Rhododendron yunnanensis (sample 2), Rhododendron chinensis (sample 3), Rhododendron grandiflora (sample 4) and Rhododendron wide cup (sample 5) were used as experimental materials. , extract the total DNA of Rhododendron alpine dry leaves with the following improved 4×CTAB method, that is, the method of the present invention. Materials such as the five different species of Rhododendron alpine and various reagents used in the examples are commercially available.

[0031] (1) Material grinding

[0032] Weigh 100 mg of Rhododendron alpine dry leaves and place them in a mortar with liquid nitrogen added in advance to grind them into powder; since the collected fresh Rhododendron alpine leaves need to be immediately ground in liquid nitrogen to prevent loss of DNA, the inventors used fresh Rhododendron alpine leaves and dried leaves as experimental materials, and compared the DNA extraction effects of 2 × CTAB, 4 × CTAB and improved 4 × CTA...

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Abstract

The invention discloses an improved method for extracting total DNA (deoxyribonucleic acid) from polysaccharide and polyphenol plant Rhododendron lapponicum. A polysaccharide removal treatment step is added to the method, dried Rhododendron lapponicum leaves are ground into powder with liquid nitrogen, then 1 mL of Buffer I is added, and most secondary metabolites such as polysaccharides, polyphenols and the like in a plant cell homogenate are washed through ice bath and centrifugation; a cell lysis solution adopts 4*CTAB (cetyltrimethylammonium bromide); cell lysis mixed liquor is not extracted directly during DNA extraction, but a supernatant obtained from separation after high-speed centrifugation of the cell lysis mixed liquor is extracted; a Rnase A reagent is added to a supernatant obtained through first extraction, the obtained mixture is treated at the room temperature for 20 min, and then second conventional extraction is performed; during washing of DNA precipitates, the arranging direction of a centrifuge tube is changed to the opposite for next washing, and the high-speed centrifugation time is prolonged to 5 min. The method has the advantages of high yield, high purity and good repeatability and can be applied to downstream experiments of molecular biology.

Description

technical field [0001] The invention relates to the technical field of plant life science, in particular to an improved high-efficiency (high yield and high purity) DNA extraction method for the polysaccharide polyphenol plant rhododendron alpine. Background technique [0002] Rhododendron L. is a world-renowned garden ornamental plant, one of the top ten famous flowers in my country and one of the eight famous flowers in Yunnan Province, China. Alpine Rhododendron is a general term for some original species of evergreen broad-leaved Rhododendron and its hybrid offspring. Because of its beautiful plant shape, large and colorful flowers, and evergreen seasons, it has gradually become a rookie in my country's New Year's Eve flowers and high-end landscaping. my country has more than 320 species of rhododendrons, accounting for 1 / 3 of the world's rhododendrons. The southwest and adjacent eastern Himalayas are the distribution centers and origins of the evergreen subgenus Rhodode...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 张露李世峰蔡艳飞解玮佳宋杰彭绿春李树发陆琳
Owner FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI
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