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Kit for rapid detection of red tide algae in natural water sample

A red tide algae and kit technology, applied in the field of molecular biology, can solve problems such as kits that do not have natural red tide water samples, and achieve the effects of good application and market requirements, simple operation steps, and easy portability.

Inactive Publication Date: 2012-03-14
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the above method has been reported to detect natural water samples, there is no report on a portable test kit that can detect natural red tide water samples.

Method used

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  • Kit for rapid detection of red tide algae in natural water sample
  • Kit for rapid detection of red tide algae in natural water sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Detection of Prorocentrum donghaiense in water samples to be tested:

[0027] Kit: The kit consists of filter membrane, reagent and sample to be tested; the reagent is 0.1mlddH 2 O. 0.3ml 95% ethanol, 2.1ml 20×SET buffer, 600μl 5×SET buffer, 400μl 1×SET buffer and 10μl FITC-labeled oligonucleotide probe solution; 2.5ml sample to be tested.

[0028] The FITC-labeled oligonucleotide probe is designed according to the specific sequence of the ITS region of Prorocentrum donghaiense, the sequence is: 5'-TCCTGATCGTCTCCTGCCT-3'.

[0029] How to use the kit:

[0030] (1) Fixation of algae cells: Add 0.1ml ddH to a 5ml sterilized centrifuge tube 2 O, 0.3ml of 95% ethanol and 2.1ml of 20×SET buffer and mix well, add 2.5ml of natural water sample to be tested collected from the red tide outbreak area in the East China Sea to an Eppendorf centrifuge tube, mix upside down, and let stand at room temperature for 30min;

[0031] (2) Filtration of algae cells: absorb the above-mentio...

Embodiment 2

[0039] Detection of H. akashiwo in water samples to be tested:

[0040] Kit: The kit consists of filter membrane, reagent and sample to be tested; the reagent is 0.05mlddH 2 O. 0.2ml 95% ethanol, 1.5ml 20×SET buffer, 700μl 5×SET buffer, 500μl 1×SET buffer and 12μl FITC-labeled oligonucleotide probe solution; 2.5ml sample to be tested.

[0041] The FITC-labeled oligonucleotide probe is designed according to the specific region of the ribosome large subunit of Heterocurvium akashiwo, and the sequence is: 5'-ACGAGCTTCCAGAATGCCAA-3'.

[0042] Instructions:

[0043] (1) Fixation of algae cells: add 0.05ml ddH to a 5ml sterilized centrifuge tube 2 O. 0.2ml is 95% ethanol and 1.5ml of 20×SET buffer and mix well, add the natural water sample collected from the East China Sea red tide outbreak area to concentrate 2-3 times, take 2.5ml of the water sample to be tested in an Eppendorf centrifuge tube, Inverted to mix, stand at room temperature for 40min;

[0044] (2) Filtration of al...

Embodiment 3

[0052] Detection of Phaeocystis in water samples to be tested:

[0053] Kit: The kit consists of filter membrane, reagent and sample to be tested; the reagent is 0.2mlddH 2 O. 0.4ml 95% ethanol, 2.5ml 20×SET buffer, 650μl 5×SET buffer, 450μl 1×SET buffer and 15μl FITC-labeled oligonucleotide probe; 1.9ml sample to be tested.

[0054] The FITC-labeled oligonucleotide probe solution is designed according to the specific region of the Phaeocystis ITS region, the sequence is: 5'-TCATCGAGACTTTGAACGCACATG-3')

[0055] (1) Fixation of algae cells: Add 0.2ml ddH to a 5ml sterilized centrifuge tube 2 O. 0.4ml of 95% ethanol by volume and 2.5ml of 20×SET buffer and mix well, add the natural water sample collected from the red tide outbreak area in the East China Sea to concentrate 2-3 times, take 1.9ml of the water sample to be tested and centrifuge in Eppendorf Tube, mix by inverting, and let stand at room temperature for 45min;

[0056] (2) Filtration of algae cells: absorb the abo...

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Abstract

The invention, relating to a molecular biology technology, particularly discloses a kit for rapid detection of red tide algae in a natural water sample and a usage method thereof. The kit comprises a filter membrane, a reagent and a sample to be detected, wherein, the reagent comprises 0.05-0.2 mL of ddH2O, 0.2-0.4 mL of 95% ethanol, 1.5-2.5 mL of 20*SET buffer, 600-700 mu l of 5*SET buffer, 400-500 mu l of 1* SET buffer and 10-15 mu l of FITC labeled oligonucleotide probe solution. The kit is convenient to carry, and can be carried to the ships for detecting red tide algae in the natural water sample at any time. Simultaneously, the kit has the advantages of simple operation, time saving, and effectiveness, and has good application and market demands.

Description

technical field [0001] The invention relates to molecular biology technology, in particular to a kit for rapidly detecting red tide algae in natural water samples. Background technique [0002] Red tide (red tide) generally refers to the phenomenon of discoloration of sea water due to the excessive reproduction of marine plankton (mainly dinoflagellates and diatoms). Strictly speaking, red tide is a natural phenomenon, but since the 1970s, there have been more and more reports about red tide. According to incomplete statistics, in 1972 and 1998, there were There are 360 ​​recorded red tides. The frequency of red tides has increased significantly, the scale of occurrence and the degree of damage have become increasingly serious, which not only affects the marine ecological environment of our country, but also causes huge losses to marine fisheries. What's more serious is that the frequent outbreaks of red tides make the species of red tide algae expand rapidly around the wo...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
Inventor 王广策张宝玉陈国福
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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