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Method for extracting deoxyribonucleic acid (DNA) from mature kenaf laminas

A technology of mature leaves and kenaf, which is applied in the field of DNA extraction of mature kenaf leaves, can solve the problems of complex steps and operations, affecting the efficiency of kenaf DNA extraction, etc., achieve good repeatability, clear agarose electrophoresis bands, and increase yield Effect

Inactive Publication Date: 2012-07-04
FUJIAN AGRI & FORESTRY UNIV
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Problems solved by technology

At present, there are few reports on the extraction method of kenaf genome DNA, and all of them use young and fresh leaves as test materials (Xu Jiantang et al., 2007; Guo Anping, 1997). There are also reports (Cao Deju, 1999; Bai Fenghu, 2007), but due to the complicated steps, the extraction efficiency of kenaf DNA is seriously affected

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  • Method for extracting deoxyribonucleic acid (DNA) from mature kenaf laminas
  • Method for extracting deoxyribonucleic acid (DNA) from mature kenaf laminas
  • Method for extracting deoxyribonucleic acid (DNA) from mature kenaf laminas

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Embodiment Construction

[0014] The specific implementation steps of the present invention are as follows:

[0015] 1.1 DNA extraction method steps of mature leaves of kenaf

[0016] Modified SDS method

[0017] (1) Weigh 1.0g of each mature leaf of the kenaf variety Fuhong 952, quickly grind it with liquid nitrogen 1-2 times until it becomes powdery, put the powder into a 10ml centrifuge tube, and add 3ml of 65°C preheated SDS extraction buffer (100 mmol / L Tris-HC1 (pH8.0), 0.5 mol / L NaC1, 20 mmol / L EDTA (pH 8.0)), add different CTAB concentrations and antioxidant combinations to each tube, 8 Different CTAB concentrations and antioxidant combinations are as follows.

[0018]

[0019] (2) After cooling to room temperature, add an equal volume of chloroform / isoamyl alcohol (24 / 1), gently invert and mix for 10 minutes; (3) Centrifuge at 1200rpm for 10 minutes; (4) Transfer the supernatant to a 10ml centrifuge tube, add 1 / 3 volume of 3 mol / L NaAc and an equal volume of -20°C pre-cooled absolute e...

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Abstract

The invention belongs to the field of plant molecular biology, and particularly relates to a method for efficiently and stably extracting deoxyribonucleic acid (DNA) from mature kenaf laminas. According to the invention, a method for extracting genome DNA from kenaf laminas by a cetyltrimethyl ammonium bromide (CTAB) method is improved and optimized on the basis of a traditional kenaf genome DNA extraction method by combining with the characteristic that the mature kenaf laminas contain polysaccharide and polyphenol. The kenaf genome DNA obtained by applying the method for efficiently and stably extracting the DNA from the mature kenaf laminas has clear strips and do not have tailings; OD260 / OD280 is 1.7-1.9; enzyme digestion is complete; the quality and the yield of the kenaf genome DNA obtained by applying the method for efficiently and stably extracting the DNA from the mature kenaf laminas are higher than that of the kenaf genome DNA obtained by applying an improved sodium dodecyl sulfonate (SDS) method. The kenaf genome DNA can be used for amplifying chloroplast DNA and mitochondria DNA. A working foundation is laid for further developing researches on genomics of kenaf and bast-fibre plants thereof and subcellular genomics.

Description

technical field [0001] The invention belongs to the field of plant molecular biology and relates to a kenaf DNA research technology, in particular to a high-quality, high-yield DNA extraction method for mature kenaf leaves. Background technique [0002] Kenaf (Kenaf) is Malvaceae (Malvaceae) Hibiscus genus ( Hibiscus ) annual bast fiber crop, which was cultivated in African Sudan in 4000 BC. As a natural fiber crop, kenaf has the characteristics of fast growth, strong stress resistance, and wide adaptability. The huge biological yield is 3- 4 times, the extremely strong carbon dioxide absorption capacity is 4-5 times that of forests, and it is an advantageous crop for developing a low-carbon economy. In addition to being traditionally used as raw materials for processing hemp rope, sacks, carpets, etc., kenaf fiber is also a good material for processing automotive interiors, panels, and hemp carbon. Antibacterial and other biological functions. [0003] With the developme...

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Application Information

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IPC IPC(8): C12N15/10
Inventor 徐建堂祁建民陶爱芬方平平林荔辉林培清陈涛池仁漫
Owner FUJIAN AGRI & FORESTRY UNIV
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