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PCR primer pair for detecting cactus X virus and applications of PCR primer pair

A cactus and primer pair technology, which is applied to the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems such as the detection method is yet to be established, the production of dragon fruit is decreased, and the lack of treatment measures, etc. Good practical value, improved detection efficiency, simple and convenient operation

Pending Publication Date: 2016-07-27
INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Viral diseases on dragon fruit have always been one of the main limiting factors of dragon fruit production and cultivation in the world, among which cactus X virus ( Cactus virus X , CVX) is an important virus that causes serious diseases of pitaya, leading to a decrease in yield and quality of pitaya
For the virus, there is still a lack of effective treatment measures. Once the virus becomes popular, it will cause heavy economic losses. At present, the main focus is on prevention.
Since cactus X virus has only been reported in China (except Taiwan) in recent years, detection methods for this virus have yet to be established

Method used

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  • PCR primer pair for detecting cactus X virus and applications of PCR primer pair
  • PCR primer pair for detecting cactus X virus and applications of PCR primer pair
  • PCR primer pair for detecting cactus X virus and applications of PCR primer pair

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Experimental program
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Effect test

Embodiment 1

[0049] The detection method of the detection of the dragon fruit on the dragon fruit provided by this embodiment, including the following steps:

[0050] (1) Extract the total RNA of the dragon fruit sample and disease sample, spare; the specific steps of the extra total RNA are as follows:

[0051] A.Take 0.2g of Dragon Fruit Sample in the mantle. After the liquid nitrogen is frozen (8 ~ 10s), grind it into powder, and then the grinding powder is used to 1.5ml of centrifugal tube;

[0052] B.Add a 2 × CTAB buffer of 1ml65 ° C to the centrifugal tube of step A, and after mixing, 65 ° C for 10 minutes;

[0053] C.Add 500 μl chloroform / isopreyol (volume ratio, 24: 1), mix 4 ℃, 12000r / min centrifuged 10min;

[0054] D.Take 800 μL to clear, add 800 μL of chloroform / isopreyol (24: 1), mix 4 ° C, 12000r / min centrifuged 10min;

[0055] E.Take 450 μL to clear, add 2 times the volume of water-free ethanol at 4 ° C, and place it for 30 minutes;

[0056] F.4 ° C, 12000r / min centrifuged 15min...

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Abstract

The invention belongs to the technical field of plant virus detection, and in particular relates to PCR primers for detecting a cactus X virus and applications of the PCR primers. The primers comprise an upstream primer CVX-CPF: 5'-CAGTCCGCCGGACCCCG-3', and a downstream primer CVX-CPR: 5'-AGACTCTGAACCTTGGAGCCCC-3'. The PCR primers provided by the invention have certain universality for detecting CVX, can be used for detecting the CVX virus on dragon fruits, and also can be used for detecting the CVX virus in cactus. According to a detection method for detecting the CVX virus on dragon fruits, the inverse transcription and PCR amplification are carried out in one step, thus the probability of cross contamination can be greatly reduced, and meanwhile, the detection efficiency is greatly improved. The practical applications show that the operation is simple and convenient, the detection sensitivity is relatively high, and about 1 pg can be detected.

Description

Technical field [0001] This application belongs to the field of plant virus detection technology, which involves a PCR primer for detecting PCR primers used for the detection of fairy palm X virus. Background technique [0002] Dragon fruit is a typical tropical fruit plant (or the genus of snake whip columns) of the cactus., Light with aroma; rich nutrition, in addition to carbohydrates and crude fibers, also contain unsaturated fatty acids, antioxidant substances, vitamin B clan, carotene, chlorophyll, mineral calcium, phosphorus, iron, etc.Low fat, low calories and other characteristics.In addition, dragon fruit, especially red meat and purple varieties, rich in anthocyanins, and water -soluble dietary fiber and plant protein rich in other fruits, which can prevent constipation, reduce blood sugar, blood lipids, and detoxify and moisturize the lungs.It is one of the best fruits in the fruit market and one of the best fruit varieties in the fruit market. [0003] Fire Dragon Fr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/701C12Q2545/113
Inventor 余乃通刘志昕龚殿王健华何凡罗志文李叶周启林
Owner INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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