YUAN Lei,LIU Yu,ZHONG Zhengchang,et al.SCoT-PCR Reaction System for Rubus biflorus Buch[J].Northern Horticulture,2017,41(03):109-111.[doi:10.11937/bfyy.201703025]
粉枝莓SCoT分子标记的PCR反应体系建立
- Title:
- SCoT-PCR Reaction System for Rubus biflorus Buch
- Keywords:
- Rubus biflorus Buch; SCoT; optimization
- 文献标志码:
- A
- 摘要:
- 以粉枝莓为试材,以改良CTAB法提取的基因组DNA为模板,从引物、dNTPs、Mg2+浓度以及退火温度4个因素,对SCoT-PCR反应体系进行了优化,以建立适合粉枝莓SCoT-PCR最佳扩增体系。结果表明:SCoT-PCR反应体系的最佳条件为dNTPs 0.125 mmol?L-1、Taq DNA聚合酶0.15 μL、引物1.0 μmol?L-1、Mg2+ 0.625 mmol?L-1、模板40 ng、反应体积20 μL、退火温度为51 ℃。该优化体系扩增的产物条带清晰,稳定性高。
- Abstract:
- Rubus biflorus Buch was used as test material,total genomic DNA was extracted by using improved CTAB method.The factors including primers,dNTPs,Mg2+ and annealing temperature were studied to establish optimized SCoT-PCR reaction system to ensure the stability of SCoT for Rubus biflorus Buch.The results showed that the optimized reaction system of SCoT-PCR was consisted as follows,in the total reaction system with amount of 20 μL,there were 0.125 mmol?L-1 dNTPs,0.15 μL Taq DNA polymerase,1.0 μmol?L-1 primers,0.625 mmol?L-1 Mg2+,40 ng template DNA,respectively.The optimal annealing temperature was 51 ℃.The findings suggested that the products′ bands were clear with high stability.
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备注/Memo
第一作者简介:袁雷(1982-),男,硕士,副教授,研究方向为天然产物。E-mail:249901708@qq.com.基金项目:西藏自治区自然科学基金资助项目(2015ZR-13-30);西藏野生特色生物资源开发平台建设资助项目(PT2015-01)。