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Position: Home > Articles > Optimization and Application of SSR-PCR Reaction System in Sun-Cured Tobacco Hubei Agricultural Sciences 2013,52 (9) 2070-2073

晒烟SSR-PCR反应体系的优化与应用

作  者:
王曼;宋长征;金爱兰;金东淳;朴世领
单  位:
吉林省海龙镇农业技术推广站;吉林省延边朝鲜族自治州农业科学院;延边大学农学院
关键词:
晒烟;SSR-PCR;优化;筛选
摘  要:
以晒烟8107为试材,对影响SSR-PCR反应体系的主要因子进行单因素优化试验,建立最适合晒烟的SSR-PCR反应体系。结果表明,20μL体系中含20 ng模板DNA、0.25μmol/L SSR引物、1.5 mmol/LMg2+、0.2 mmol/L dNTPs、1.5 U Taq酶,最适退火温度为56.7℃。筛选出16对扩增条带清晰、多态性较强的引物,并对12个晒烟品种进行扩增,初步分析了SSR标记应用于晒烟遗传多样性与亲缘关系研究的潜力。
译  名:
Optimization and Application of SSR-PCR Reaction System in Sun-Cured Tobacco
作  者:
WANG Man1,2,SONG Chang-zheng3,JIN Ai-lan2,JIN Dong-chun1,PIAO Shi-ling1(1.Agricultural College,Yanbian University,Yanji 133002,Jilin,China;2.Academy of Agricultural Sciences of Yanbian,Longjing 133400,Jilin,China;3.Hailong Town Agricultural Technology Extension Station in Jilin Province,Hailong 135007,Jilin,China)
关键词:
sun-cured tobacco;SSR-PCR;optimization;screening
摘  要:
The main factors of SSR-PCR reaction system were analyzed by univariate optimization experiments based on the genomic of sun-cured tobacco 8107,and the stable SSR-PCR system was established for sun-cured tobacco.The results showed that 20 μL reaction solution contained 20 ng DNA,0.25 μmol / L SSR primers,1.5 mmol / L Mg2+,0.2 mmol / L dNTPs,1.5 U Taq polymerase,and the optimal annealing temperature of the primer was 56.7 ℃.16 pairs of primers were picked out and used to amplify 12 copies sun-cured tobacco,the potential of SSR markers that used in the relationship between suncured tobacoo germplasm genetic diversity and phylogenetic was initially analyzed.

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