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Molecularly specific labeling primers and detection method for mountain rice Bailinggu No.2 and No.19

A marker primer, specific technology, applied in the direction of DNA/RNA fragments, recombinant DNA technology, etc., can solve the problem of unsatisfactory identification of rice varieties, and achieve the effect of simple method

Inactive Publication Date: 2015-05-13
ZHEJIANG FORESTRY ACAD
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Problems solved by technology

Some PCR-based molecular marker technologies such as RAPD (Random Amplified Polymorphic DNA, Random Amplified Polymorphic DNA), AFLP (Amplified Fragment Length Polymorphism, Amplified Fragment Length Polymorphism) have been used in various species classification and identification, but these markers are not ideal for the identification of mountain rice varieties

Method used

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  • Molecularly specific labeling primers and detection method for mountain rice Bailinggu No.2 and No.19
  • Molecularly specific labeling primers and detection method for mountain rice Bailinggu No.2 and No.19
  • Molecularly specific labeling primers and detection method for mountain rice Bailinggu No.2 and No.19

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Embodiment 1

[0033] (1) Genomic DNA extraction: Genomic DNA was extracted using the SDS-CTAB method. The purified genomic DNA is first qualitatively detected by 1.5% agarose gel electrophoresis, and then quantitatively detected by a DNA / RNA ultraviolet spectrophotometer. DNA extracts were stored in a -20°C refrigerator for later use.

[0034] (2) Design specific PCR amplification primers. The sequence of the primer pair is the upstream primer 5'-AGCGGCCGCATCCTGAGAGA-3' and the downstream primer 5'-AGCGGCCGCATCCTGAGAG-3', which were synthesized by Shanghai Sangon Bioengineering Technology Co., Ltd.

[0035] (3) PCR amplification of SCAR molecular markers:

[0036] The composition of each 20 μL PCR amplification system is as follows:

[0037]

[0038]

[0039] The amplification reaction was carried out on a TC-XP type amplification instrument.

[0040] The PCR amplification conditions are as follows:

[0041] Pre-denaturation at 94°C for 6 minutes; denaturation at 92°C for 40 s, an...

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Abstract

The invention relates to a pair of high-specificity molecularly specific labeling primers for mountain rice Bailinggu No.2 and No.19 which have high specificity and a method capable of rapidly detecting the mountain rice Bailinggu No.2 and No.19. The sequences of the primers are as follows: the sequence of the upstream primer is 5'-AGCGGCCGCATCCTGAGAGA-3' and the sequence of the downstream primer is 5'-AGCGGCCGCATCCTGAGAG-3'. By using the molecularly specific labeling primers, the early identification of mountain rice Bailinggu No.2 and No.19 can be rapidly performed, and the method is simple, fast and accurate and is an irreplaceable molecular tool for distinguishing mountain rice varieties in apparent characteristics.

Description

(1) Technical field [0001] The invention relates to molecular specific marker primers of mountain rice Bailinggu No. 2 and Bailinggu No. 19 and identification methods thereof. (2) Background technology [0002] Mountain rice has a long history of cultivation in my country and is widely distributed, especially in ethnic minority areas such as Yunnan and Guizhou. It mainly adopts the nomadic slash-and-burn production method. Long-term natural selection and artificial cultivation have formed my country's rich mountain rice germplasm resources. In recent years, with the development of the under-forest economy, the construction of various under-forest economic development models with large scale, good benefits and strong driving force has become the main goal of the country for the development of the under-forest economy. However, for a long time, the production and sales of mountain rice in China's main rice producing areas have been relatively chaotic, and the phenomenon of "d...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895
Inventor 王丽玲刘本同秦玉川钱华王衍彬
Owner ZHEJIANG FORESTRY ACAD
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