高丹草SRAP-PCR反应体系优化及验证

doi:10.12190/j.issn.2096-1197.2022.01.13

Journal of Northern Agriculture ›› 2022, Vol. 50 ›› Issue (1): 93-100.doi: 10.12190/j.issn.2096-1197.2022.01.13

• Plant protection・Aquaculture • Previous Articles Next Articles

Optimization and verification of a simple sequence-related amplified polymorphism（SRAP）-PCR system in Sorghum-Sudan hybrid grass

SHI Yue¹, LIU Shuancheng¹, FANG Yongyu², DING Haijun², LYU Ersuo², LU Yuhong³

1. College of Life Science and Technology,Jining Normal University,Jining 012000,China;
2. Institute of Grassland,Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences,Hohhot 010031,China;
3. Agricultural Technolongy Extension Station of Sijiazi Town in Aohan Banner,Sijiazi 024300,China

Received:2022-01-11 Online:2022-02-20 Published:2022-04-11

Abstract

Abstract: 【Objective】To find the optimal Sorghum-Sudan hybrid grass SRAP-PCR reaction system.【Methods】The five components（dNTP,template DNA concentration,primers,Taq DNA polymerase,and Mg²⁺）were optimized using single-factor and orthogonal-design approaches. The optimized SRAP-PCR reaction system was verified.【Results】The best SRAP-PCR system for Sorghum-Sudan hybrid grass contained 275 μmol/L dNTP,3.0 ng/μL template DNA,1.1 μmol/L for each primer,1.00 U Taq DNA polymerase,and 2.25 mmol/L Mg²⁺,making up to 20 μL with ddH₂O. 【Conclusion】The optimized SRAP-PCR reaction system amplified many bands,with abundant polymorphism and good repeatability.

Key words: Sorghum-Sudan hybrid grass, SRAP-PCR, Orthogonal design, System optimization, Verification

CLC Number:

S544.9

SHI Yue, LIU Shuancheng, FANG Yongyu, DING Haijun, LYU Ersuo, LU Yuhong. Optimization and verification of a simple sequence-related amplified polymorphism（SRAP）-PCR system in Sorghum-Sudan hybrid grass[J].Journal of Northern Agriculture, 2022, 50(1): 93-100.

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Optimization and verification of a simple sequence-related amplified polymorphism（SRAP）-PCR system in Sorghum-Sudan hybrid grass

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