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Method for rapidly authenticating green tide algae enteromorpha

A prolifera and algae technology, which is applied in the field of rapid identification of green tide algae prolifera, can solve the problems of complex and time-consuming identification of prolifera.

Inactive Publication Date: 2011-01-12
OCEAN UNIV OF CHINA
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Problems solved by technology

[0008] The invention provides a method for quickly identifying the green tide algae Enteromorpha, which can solve the problems of complicated and time-consuming identification of Enteromorpha existing in the prior art

Method used

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  • Method for rapidly authenticating green tide algae enteromorpha
  • Method for rapidly authenticating green tide algae enteromorpha
  • Method for rapidly authenticating green tide algae enteromorpha

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Embodiment Construction

[0059] The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.

[0060] 1. DNA extraction from green algae

[0061] 1. Take fresh green algae strains (standard samples BD, FZ and samples to be tested), wash them three times with distilled water, blot the surface moisture with absorbent paper, weigh 0.3g and grind in liquid nitrogen with a mortar and pestle into a fine powder, add 3ml CTAB extract (100mM Tris, 1.4M NaCl, 20mM EDTA, pH 8.0, 2% CTAB, add 2-mercaptoethanol before use to make the final concentration 0.3%) and 45mg PVP, invert and mix well Digest in a water bath at 60°C for 1-2 hours, shaking continuously during the period;

[0062] 2. Take the digested mixture out of the water bath, cool it at room temperature for about 5 minutes, add an equal volume of chloroform: isoamyl alcohol (24:1) and mix it upside down to form an emulsion, then centrifuge at 6000 rpm for 15 minutes at room temp...

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Abstract

The invention discloses a method for rapidly authenticating green tide algae enteromorpha. The method fulfills the aim of rapidly authenticating enteromorpha by obtaining a ribosome RNA transcription interval spacer sequence ITS and 5s ribosome RNA serial repeat sequence interval spacer sequence 5SS by sample DNA extraction and PCR amplification by utilizing nuclear acid hybridization technology and using two DNA double-chain probes, pointing a PCR product on a nylon membrane with positive chargers after performing modification treatment by adopting the nuclear acid hybridization technology, and hybridizing the PCR product with a specific probe to develop, wherein the sample of which the ITS and the 5SS have positive signals are the enteromorpha (Ulva prolifera). The method can authenticate in simple and crude in experiment settings in the open and has high authentication speed.

Description

technical field [0001] The invention relates to a method for rapidly identifying green tide algae Enteromorpha (Ulva prolifera) through nucleic acid hybridization technology, and specifically relates to green algae DNA extraction technology, PCR amplification technology, and digoxin-labeled probes technology, nucleic acid hybridization and color development technology, and finally completed the identification of Enteromorpha. Background technique [0002] Green tide (Green tide) is an ecological anomaly that generally occurs in coastal countries around the world. It has been recorded in Europe, Asia, North America and Australia, and mainly occurs in estuaries, inner bays, lagoons and densely urbanized coasts. The main biological species that cause the green tide are Ulva sp. and Enteromorpha sp. of the Ulvaceae family. At present, they belong to the genus Ulva. Most of them are Ulva, and they also have bristles. Algal blooms (Cladophora sp.) have been reported. The green t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 茅云翔王健孔凡娜隋正红
Owner OCEAN UNIV OF CHINA
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