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Skeletonema costatum PCNA gene detecting method

A gene detection, SC-FQ-PCNA technology, applied in the field of detection of planktonic diatoms, can solve the problem that the basic physiological and ecological parameters such as the growth rate of planktonic algae cannot be measured, and achieve the stability of RNA extraction and reverse transcription system, good stability effect

Inactive Publication Date: 2008-02-27
OCEAN UNIV OF CHINA
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Problems solved by technology

[0013] The technical problem to be solved by the present invention is to provide a method for detecting the PCNA gene of Skeletons costa, which is used for estimating the productivity of planktonic algae, studying the change of planktonic algae population, constructing an ecological dynamics model and even predicting red tides, and overcoming the problem of single species in the prior art. The defect that the basic physiological and ecological parameters such as the growth rate of planktonic algae cannot be measured

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  • Skeletonema costatum PCNA gene detecting method
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Embodiment Construction

[0054] In the embodiment of the present invention, the culture process of Skeletonemacostatum is: Skeletonemacostatum (Skeletonemacostatum) is cultivated to logarithmic growth phase with F / 2 medium in the laboratory, culture conditions: light / dark cycle is 12h / 12h , the light intensity is 4000Lux, and the culture temperature is 22-25°C.

[0055] Collect 5 copies of Skeletalum costanum (cell density 4.1×107 cells / mL) cultured for one week, about 3.8×10 7 cells (the weight of the algae concentrate is about 150mg). Use the following RNA extraction methods to extract:

[0056] (1) Collect algae cells and grind them into fine powder with a mortar under the condition of continuously adding liquid nitrogen;

[0057] (2) Add the grinding liquid to 1mL TRIZOL reagent, and homogenize at room temperature for 5min;

[0058] (3) Add 200 μL of chloroform, shake vigorously for 15 seconds, and incubate at room temperature for 3 minutes;

[0059] (4) Centrifuge at 12000rpm for 15min at 4°C...

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Abstract

The present invention discloses a skeletonema costatum PCNA gene detection method. Said method includes the following steps: (1), extracting RNA of skeletonema costatum; (2), separating and cloning partial sequence of skeletonema costatum PCNA gene, utilizing 3'RACE to make the obtained skeletonema costatum PCNA sequence implement 3' end extension so as to obtain PCNA sequence with 750-850bp; and (3), according to the obtained PCNA sequence with 750-850bp constructing standard curve and utilizing said standard curve to detect the skeletonema costatum PCNA gene to be detected.

Description

technical field [0001] The invention relates to a method for detecting planktonic diatoms, in particular to a method for detecting the PCNA gene of Sclerostetes costa. Background technique [0002] S. costatum is a long chain of multiple cells, the cells are lens-shaped cylindrical cells with a diameter of 6-7 microns, 8-30 spines on the shell margin, and the intercellular space is generally larger than the length of the cells. Chromosomes are usually 2, located on the shell surface. The nucleus is in the center. [0003] Scutellaria costatum is an excellent bait for marine secondary producers such as oysters. On the other hand, Sclerotina mesocostalis is a common red tide organism in the coastal waters of my country. In economical seaweed breeding areas, it often competes with kelp and laver for nutrients. A large number of blooms of this species can cause economical algae to change color, or even rot and lose food and water. commercial value. Before and after the red ti...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 于志刚何闪英赵丽媛孙静米铁柱甄毓
Owner OCEAN UNIV OF CHINA
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