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The primer set and its products and detection methods used to identify conventional cotton China Cotton Research Institute 99001

A detection method and primer set technology, which can be used in biochemical equipment and methods, microbial determination/inspection, recombinant DNA technology, etc. The impact of the test is small and easy to observe.

Active Publication Date: 2020-07-10
INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the field identification work in the past, traditional breeders often rely on rich breeding experience to conduct field identification of conventional cotton varieties according to the biological characteristics of cotton. Although this method is effective, it is time-consuming and laborious, and it is easily affected by the external environment and human subjective. factors, not suitable for young breeders

Method used

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  • The primer set and its products and detection methods used to identify conventional cotton China Cotton Research Institute 99001
  • The primer set and its products and detection methods used to identify conventional cotton China Cotton Research Institute 99001
  • The primer set and its products and detection methods used to identify conventional cotton China Cotton Research Institute 99001

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] 1. Cotton material

[0078] Seeds of the following cotton varieties: Zhongmian Institute 99001, Upland cotton genetic standard line TM-1, Zhongmian Institute No. 10, Lumianyan No. 28, Shumian No. 2, Chuanmian 239, Emian No. 18, Simian No. 2 , Zhongmian Institute 12, Zhongmian Institute 49 and Xinlu Zhong No. 69.

[0079] 2. DNA extraction

[0080] Take the seeds of Zhongmian Institute 99001 and other 10 materials as the control, remove the shells, and use the SDS method [4] Genomic DNA was extracted and diluted to a final concentration of 50ng / μL to obtain the template to be detected.

[0081] 3. PCR amplification

[0082] The genome extracted from the above materials was used as a template to be detected, and PCR amplification was performed with 184 pairs of primers (minimum 7 pairs of primers on each chromosome on average) designed for 26 chromosomes of upland cotton. The PCR reaction system was 10 μL, the amplification mixture system was shown in Table 1, and the...

Embodiment 2

[0121] For the conventional cotton Zhongmian Institute 99001 and other cotton varieties, the method in Example 1 was repeatedly verified, and the patterns consistent with Example 1 could be obtained. It shows that the primer pair provided by the present invention can stably distinguish the conventional cotton Zhonghuasuo 99001 from other varieties.

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Abstract

The invention relates to the field of molecular detection, in particular to a primer set used for identifying conventional cotton CRI99001, a product of the primer set and a detection method. The primer set comprises any one or more of the following primer pairs: NAU1233 primer pair, DPL0308 primer pair, NAU3468 primer pair, NAU5467 primer pair, DPL0090 primer pair, DC40150 primer pair, BNL1154 primer pair, and NAU1103 primer pair. A total of 184 pairs of primers are designed, and PCR amplification is carried out on detection substances, wherein the detection substances comprise the conventional cotton CRI99001 and 10 cotton varieties, and a specific primer pair in CRI9901 varieties is obtained through comparison and screening. The primer pair has strong specificity, and the detection result is stable and reliable.

Description

technical field [0001] The invention relates to the field of molecular detection, in particular, to a primer set for identifying the conventional Cotton Institute 99001, a product thereof and a detection method. Background technique [0002] Cotton is an important economic crop in the world, which has an important influence on the economic development of our country and the world. In recent years, China's cotton planting area has long been at 500×10 4 hm 2 Wandering left and right, a "three-legged stand" pattern was formed in the northwest inland, the Yellow River Basin, and the Yangtze River Basin. The production of conventional and hybrid varieties in the three major cotton regions has been widely promoted. Hybrids have been valued by breeders for their heterosis, but as labor costs are getting higher and higher, it takes a lot of manpower and material resources, resulting in a decline in seed production. In addition, hybrid cotton has encountered many difficulties in t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2565/125
Inventor 赵俊杰任中英李威杨代刚裴小雨刘艳改贺昆仑张飞马雄风周晓箭张文生
Owner INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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