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Chromosome preparation method for enteromorpha sporophyte

A technology for spores and chromosomes of Enterobacter prolifera, which is applied in the field of cytogenetics, can solve the problems of small chromosomes and difficult preparation, and achieves the effects of simple method, simple operation and convenient acquisition.

Inactive Publication Date: 2018-05-08
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because the chromosomes of Enteromorpha sporophytes are very small, it is generally difficult to prepare, and there is no report on the chromosome preparation method of Enteromorpha sporophytes in China.

Method used

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  • Chromosome preparation method for enteromorpha sporophyte
  • Chromosome preparation method for enteromorpha sporophyte

Examples

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Embodiment 1

[0029] A method for preparing the somatic chromosome of Enteromorpha sporophyte, comprising the following steps:

[0030] (1) Material collection and pretreatment: Take a small amount of algae in good condition and put them into a 1.5mL enzyme-removing centrifuge tube, wash them twice with sterilized seawater, and centrifuge at 12000rpm for 2min to remove excess water.

[0031] (2) Colchicine treatment: add colchicine with a mass concentration of 0.2% to the pretreated algae in step (1), and place in a refrigerator at 4° C. for 6 hours.

[0032] (3) Fixation: centrifuge the material treated in step (2) to take the precipitate, add glacial acetic acid: Carnot's fixative with a volume ratio of 1:3, shake and shake well, and place it in a refrigerator at 4°C for no less than 24 hours .

[0033] (4) Enzymolysis of Enteromorpha somatic cells: wash the fixed substance in step (3) with sterilized deionized water for 3 times, centrifuge at 12000 rpm for 2 minutes, take the precipitat...

Embodiment 2

[0039] A method for preparing chromosomes of Enteromorpha sporozoites, comprising the following steps:

[0040] (1) Material collection and pretreatment: Take a small amount of algal bodies in good condition and put them into a 2mL enzyme-removing centrifuge tube, wash them with sterilized seawater for 3 times, and centrifuge at 12000rpm for 2min to remove excess water.

[0041] (2) Colchicine treatment: add colchicine with a mass concentration of 0.2% to the algae pretreated in step (1), and place in a refrigerator at 4° C. for 7 hours.

[0042] (3) Fixation: centrifuge the material treated in step (2) to take the precipitate, add glacial acetic acid: Carnot's fixative with a volume ratio of 1:3, shake and shake well, and place it in a refrigerator at 4°C for no less than 24 hours .

[0043] (4) Enzymolysis of Enteromorpha somatic cells: Wash the fixed substance in step (3) with sterilized deionized water for 4 times, centrifuge at 12000rpm for 2min, take the precipitated al...

Embodiment 3

[0048] A method for preparing chromosomes of Enteromorpha sporozoites, comprising the following steps:

[0049] (1) Material collection and pretreatment: Take a small amount of algae in good condition and put them into a 1.5mL enzyme-removing centrifuge tube, wash them twice with sterilized seawater, and centrifuge at 12000rpm for 2min to remove excess water.

[0050](2) Colchicine treatment: add colchicine with a mass concentration of 0.2% to the pretreated algae in step (1), and place in a refrigerator at 4°C for 8 hours;

[0051] (3) Fixation: centrifuge the material treated in step (2) to take the precipitate, add glacial acetic acid: Carnot's fixative with a volume ratio of 1:3, shake and shake well, and place it in a refrigerator at 4°C for no less than 24 hours .

[0052] (4) Enzymolysis of Enteromorpha somatic cells: wash the fixed substance in step (3) with sterilized deionized water for 5 times, centrifuge at 12000rpm for 2min, take the precipitated algae, add appro...

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Abstract

The invention discloses a chromosome preparation method for enteromorpha sporophyte. The chromosome preparation method comprises the following steps: taking materials, dewatering, colchicine pretreatment, immobilizing, performing enzymolysis by a mixed enzyme solution, stopping enzymolysis by 70% of ethanol, high-level dropping and DAPI (4,6-Diamidino-2-Phenylindole); treating enteromorpha sporophyte cells by colchicines so as to be immobilized at metaphase in cell division, performing high-level dropping after mixed enzyme solution treatment, completely removing cell walls, cell membranes, intracellular polysaccharides and other substances of the enteromorpha sporophyte, thereby realizing cell dispersing uniformity and excellent chromosome spread. The background is clear, and relatively more cells are divided at the metaphase. The chromosome of the enteromorpha sporophyte prepared by the method is a 1-2-micron short rod-like small chromosome, 2n is equal to 18, and subsequent karyotype analysis of the enteromorpha sporophyte is facilitated.

Description

technical field [0001] The invention belongs to the field of cytogenetics, in particular to a chromosome preparation method of enteromorpha sporophyte. Background technique [0002] Green tide is a harmful ecological phenomenon caused by the explosive growth of certain green algae. It has developed into a global environmental problem and is attracting more and more attention. The outbreak of green tide causes a large amount of green algae to accumulate on the coast, which seriously affects the offshore and coastal landscape, and has a negative effect on the tourism economy such as tourism and water sports. In addition, a large amount of green algae biomass will cause a series of secondary disasters during the outbreak of green tides, endangering the ecological balance of the coastal environment and marine communities. Since the Yellow Sea green tide first occurred in 2007, it has been a large-scale outbreak for 10 consecutive years, and Ulva prolifera was identified as the ...

Claims

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Application Information

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IPC IPC(8): G01N1/28G01N1/30G01N21/64
CPCG01N1/2813G01N1/30G01N21/6458
Inventor 赵晓惠张建恒陈冉霍元子蔡春尔何培民
Owner SHANGHAI OCEAN UNIV