Molecular identification method for China rana rugulosa and Thailand rana rugulosa

A molecular identification and tiger frog technology, applied in the biological field, to achieve reliable detection results, advanced and reasonable implementation steps, and advanced and reasonable technical means

Inactive Publication Date: 2014-03-26
ZHEJIANG NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented method uses special techniques called DNA sequencing or polymerase chain reaction (PCR) to accurately detect Japanese tapirflies on their host fishes. By comparing these sequences with existing methods like histological analysis, we found they have strong evidence against certain species such as China's tick-spoons. These experiments showed that this technique could lead us towards understanding how pests live inside caterpillars living around Japan.

Problems solved by technology

This patents describes how tickets like Turtles can hide their appearance when they live outdoors without being seen by humans. These hidden indications may include dirtiness (such as pimples) around certain areas such as those near eyesightedness or behind joint pain syndrome. Additionally, these signs could indicate diseases caused by bacteria called Bacillus cerevisiae which causes swelling inside fishing nerves. Therefore, identifying this type of seafood requires quick and precise techniques to help authorities detect any potential threatening organisms.

Method used

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  • Molecular identification method for China rana rugulosa and Thailand rana rugulosa
  • Molecular identification method for China rana rugulosa and Thailand rana rugulosa
  • Molecular identification method for China rana rugulosa and Thailand rana rugulosa

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Embodiment

[0051] The Chinese tiger frogs collected in Yunnan, Zhejiang, and Hubei, and the Thai tiger frogs, American frogs, bullfrogs, spiny-bellied frogs, spiny-breasted frogs, black-spotted frogs, and Chinese forest frogs collected in Zhejiang were used as identification objects .

[0052] Take 7 samples of Chinese tiger frog, 6 samples of Thai tiger frog, 1 sample of American frog, 1 sample of Chinese forest frog, and 2 samples of bullfrog, spiny-bellied frog, spiny-breasted frog and black-spotted frog. The specific operation steps are as follows:

[0053] 1) Sample DNA extraction:

[0054] ① Take a few pairs of scissors and tweezers, soak them in absolute ethanol, take samples of each frog, rinse the toes with sterile water, and dry the surface water with filter paper;

[0055] ② Burn the scissors on the alcohol lamp, clamp the toes with tweezers, cut the toes of each sample to 3mm after cooling, and place them in a 1.5ml centrifuge tube;

[0056] ③ Wash the toes of the frog in ...

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Abstract

In order to solve the identification problem of China rana rugulosa and Thailand rana rugulosa which have exactly-like appearance during actual protection of China rana rugulosa, the invention provides a molecular identification method for China rana rugulosa and Thailand rana rugulosa. The identification method is based on PCR technology, helps to realize rapid identification on China rana rugulosa and Thailand rana rugulosa by designing a specific primer for amplification, and specifically comprises the steps: 1) extracting DNA of a sample; 2) detecting sample DNA; 3) performing PCR amplification by employing four pairs of specific primers; and 4) performing identification on the specific amplified product. According to the method, by designing the four pairs of primers related to different gene positions, China rana rugulosa and Thailand rana rugulosa can be identified by PCR amplification, the primers are strong in specificity, the agarose gel electrophoresis characteristic spectral band is high in stability and has no regional difference, and the method can help to obtain reliable beneficial effect during practical application.

Description

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Claims

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Application Information

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Owner ZHEJIANG NORMAL UNIVERSITY
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