Method for simultaneously extracting high quality DNA/RNA of kelp gametophyte

A gametophyte, high-quality technology, applied in the field of molecular biology, can solve the problems of liquid nitrogen grinding, small DNA fragments, large amount of materials, etc., and achieve the effects of convenient material acquisition, avoiding interference and saving costs.

Inactive Publication Date: 2015-03-11
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The invention provides a simultaneous preparation method of kelp gametocyte DNA/RNA, which can solve the problems existing in the prior art,

Method used

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  • Method for simultaneously extracting high quality DNA/RNA of kelp gametophyte
  • Method for simultaneously extracting high quality DNA/RNA of kelp gametophyte
  • Method for simultaneously extracting high quality DNA/RNA of kelp gametophyte

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Embodiment 1

[0029] A method for simultaneously extracting high-quality Laminaria gametophyte DNA / RNA, which includes material selection, multiple grinding and cracking of materials, extraction, precipitation and dissolution, and purification of DNA / RNA; the specific operations are as follows:

[0030] Take fresh kelp gametophytes and wash them in filtered seawater, measure the fluorescence parameter Fv / Fm value of gametophyte chlorophyll, and select the gametophytes with Fv / Fm value between 0.3-0.6.

[0031]1g gametocytes are placed in mortar, add and contain polyvinylpyrrolidone (PVP) and dithiothreitol (DTT) CTAB lysis buffer 10ml, the final concentration of PVP and DTT in CTAB lysis buffer is respectively 2% (mass ratio) and 50mM. Grind until the tissue is dissolved; divide the tissue lysate into six 2ml Ep tubes, add two 3mm steel balls to the Ep tubes, vortex, and grind for 20min with a tissue grinder at 10°C; then centrifuge (45sec , 8100×g), collected the supernatant in a 50ml cen...

Embodiment 2

[0041] In this embodiment, the conventional liquid nitrogen grinding method is used to extract the DNA in the gametophyte, and the remaining steps are completely the same as in Example 2, and finally the DNA solution is obtained, and the quality of the nucleic acid sample is detected by electrophoresis on a 1% agarose gel, as shown in FIG. Figure 5 .

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Abstract

The invention discloses a method for simultaneously extracting high quality DNA/RNA of kelp gametophyte, and belongs to the field of molecular biology. The method comprises steps of material selection, material grinding and cracking for several times, extraction, precipitation, dissolution, and DNA/RNA purification. The provided method can solve the problems that in the prior art the material consumption is huge, the materials should be grinded by liquid nitrogen, the obtained DNA/RNA amount is little, the DNA section is small, and the polysaccharide pollution rate is high. The invention aims to provide a simple and practical preparation method, which is suitable for being carried out in a lab, of kelp gametophyte DNA/RNA.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular to a method for simultaneously extracting high-quality laminar gametophyte DNA / RNA. Background technique [0002] Kelp is a plant native to the coasts of Japan, Korea, and the Far East of the former Soviet Union. Kelp was introduced to my country for the first time in 1927 and has gradually adapted to the marine environment of our country. Kelp has become an important economic seaweed in the world. It has a wide range of uses and is widely used in medicine, food, chemical industry and so on. As an excellent vegetable, most of the kelp produced in our country is used as food. In addition to eating kelp in my country, a large part of it is used as industrial raw materials to extract products such as iodine, alginate, and mannitol. Alginate is widely used in textile, printing and dyeing, medicine and food industries. Its derivatives are also important industrial raw materials. In r...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 叶乃好张晓雯苗昱范晓徐东
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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