Method for extracting nucleic acid of endangered semi-mangrove plant Hernandia nymphaeifolia

A technology of tung nucleic acid and lotus leaf, which is applied in the field of molecular biology, can solve the problems of viscous supernatant, unclean polysaccharide treatment, and wire drawing of pipette tips, etc., and achieve the effect of reducing the loss of nucleic acid
CN113136384AActive Publication Date: 2021-07-20HAINAN NORMAL UNIV
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
HAINAN NORMAL UNIV
Publication Date
2021-07-20

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Abstract

The invention belongs to the aspect of molecular biology, and particularly relates to a method for extracting nucleic acid of endangered semi-mangrove plant Hernandia nymphaeifolia. The method comprises the following steps: cracking cells by using an improved lysis solution, performing extraction by using a mixed solution of Tris balanced phenol-chloroform-isoamyl alcohol in a certain proportion, and performing extraction by using Tris balanced phenol with the same volume in order to more thoroughly remove glycoprotein; and precipitating and purifying the DNA by using sodium acetate-isopropanol with improved volume, so as to obtain the high-quality genome DNA. The DNA extracted by the method provided by the invention is high in concentration and purity, is not easy to degrade, the damage of superoxide anion free radicals or active oxygen to the DNA is prevented, and a foundation is laid for subsequent molecular biology experiments such as PCR amplification, molecular marker technology development, genome library construction and the like.
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Description

technical field

[0001] The invention belongs to the field of molecular biology, and in particular relates to a method for extracting nucleic acid of the endangered semi-mangrove plant Palismanthus japonicus. Background technique

[0002] Hernandia nymphaeifolia (J.Presl) Kubitzki is an evergreen tree belonging to the nymphaeifolia family and the nymphaeifolia genus. Hernandia nymphaeifolia is rich in polysaccharides and polyphenols and has special ingredients. The extract is extremely viscous and easy to extract. Browning, so nucleic acid extraction is more difficult.

[0003] Traditional SDS method

[0004] (1) Weigh 0.2g sample and grind it into powder with liquid nitrogen, put it in a 1.5ml centrifuge tube, add 800μL SDS lysate (100mmol / L Tris-HCl, pH8.0, 50mmol / L EDTA , 500mmol / L NaCl, 2% β-mercaptoethanol (v / v), 5% SDS (m / v)), put it in a 65°C water bath for 20min.

[0005] (2) Centrifuge at 4°C, 12 000rpm for 10min. Transfer the supernatant into a new 1.5ml centrifu...

Claims

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