[1]王玉,赵兰勇,申宏伟,等.玫瑰SCoT-PCR反应体系优化及引物筛选[J].北方园艺,2014,38(23):91-96.
　WANG Yu,ZHAO Lan-yong,SHEN Hong-wei,et al.Optimization for SCoT-PCR System and Primer Selection of Rosa rugosa[J].Northern Horticulture,2014,38(23):91-96.

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玫瑰SCoT-PCR反应体系优化及引物筛选

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 第38卷期数: 2014年23期页码: 91-96 栏目: 出版日期: 2014-12-15

Title:: Optimization for SCoT-PCR System and Primer Selection of Rosa rugosa

文章编号:: 1001-0009（2014）23-0091-06

作者:: 王玉¹; 赵兰勇¹; 申宏伟²; 齐宇¹; 石良红¹; 吴迪¹; 1.山东农业大学林学院，山东泰安 271018；
2.日照市园林绿化养护中心，山东日照 276800

Author(s):: WANG Yu¹; ZHAO Lan-yong¹; SHEN Hong-wei²; QI Yu¹; SHI Liang-hong¹; WU Di¹; 1.College of Forestry,Shandong Agricultural University,Tai’an,Shandong 271018;
2.Garden Virescence and Maintenance Center of Rizhao,Rizhao,Shandong 276800

关键词:: 玫瑰; SCoT-PCR; 正交设计; 单因素试验; 体系优化; 引物筛选

Keywords:: Rosa rugosa; SCoT-PCR; orthogonal design; single factor experiment; system optimization; primer screening

分类号:: S 685.12

文献标志码:: A

摘要:: 以玫瑰为试材，采用L₁₆（4⁵）正交设计和单因素试验2种方法，研究模板DNA、Mg²⁺、dNTPs、引物和Taq酶5个因素对玫瑰SCoT-PCR反应体系的影响，建立最优化的反应体系并筛选合适引物。结果表明：模板DNA浓度为1.50 ng/μL，Mg²⁺浓度为2.00 mmol/L，dNTPs浓度为0.35 mmol/L，引物浓度为0.70 μmol/L，Taq 酶用量为0.50 U时，可建立玫瑰SCoT-PCR最佳反应体系，并筛选出20条扩增条带清晰、多态性丰富的SCoT引物。反应体系的优化及引物的筛选,为日后利用SCoT分子标记技术对玫瑰进行相关研究提供理论依据和技术支持。

Abstract:: Taking Rosa rugosa as material，L₁₆(4⁵) orthogonal design test and single factor experiment were applied for optimizating five factors (DNA template,Mg²⁺,dNTPs,primer and TaqDNA polymerase) in the SCoT-PCR reaction system of Rosa rugosa,and primers were screened in order to establish the best reaction system of Rosa rugosa.The results showed that the best SCoT-PCR system for Rosa rugosa included 1.5 ng/μL DNA，2.00 mmol/L Mg²⁺，0.35 mmol/L dNTPs，0.70 μmol/L primer,and 0.50 U Taq DNA polymerase and 20 primers were screened which had abundant and clear polymorphism bands.The optimized SCoT-PCR system and the screeTaking Rosa rugosa as material，L₁₆(4⁵) orthogonal design test and single factor experiment were applied for optimizating five factors (DNA template,Mg²⁺,dNTPs,primer and Taq DNA polymerase) in the SCoT-PCR reaction system of Rosa rugosa,and primers were screened in order to establish the best reaction system of Rosa rugosa.The results showed that the best SCoT-PCR system for Rosa rugosa included 1.5 ng/μL DNA，2.00 mmol/L Mg²⁺，0.35 mmol/L dNTPs，0.70 μmol/L primer,and 0.50 U TaqDNA polymerase and 20 primers were screened which had abundant and clear polymorphism bands.The optimized SCoT-PCR system and the screened primers provided a theoretical basis and technical support for the further study on Rosa rugosa with SCoT-PCR.ned primers provided a theoretical basis and technical support for the further study on Rosa rugosa with SCoT-PCR

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备注/Memo

第一作者简介：王玉（1990），女，山东淄博人，硕士研究生，研究方向为园林植物造景。E-mail:532836481@qq.com.

责任作者：赵兰勇（1960-），男，山东临朐人，硕士，教授，博士生导师，研究方向为园林植物种质资源与花卉栽培及园林景观评价及园林植物造景。E-mail:sdzly369@163.com.

基金项目：山东省农业良种工程资助项目（鲁科农字(2009)103号）。

更新日期/Last Update: 2014-12-23