MENG Ling-jun,LI Zhu-gang,HAN Jun-yan,et al.Optimization for SSR-PCR Technology System in Cucurbita moschata[J].Northern Horticulture,2013,37(10):103-106.
南瓜SSR-PCR反应体系的优化
- Title:
- Optimization for SSR-PCR Technology System in Cucurbita moschata
- 文章编号:
- 1001-0009(2013)10-0103-04
- 分类号:
- S 642.1
- 文献标志码:
- A
- 摘要:
- 以3个矮生南瓜自交系和2个蔓生南瓜自交系为试材，利用Primer 5软件开发南瓜植物中具有特异性的SSR引物，并通过正交实验和梯度PCR试验对影响南瓜SSR-PCR结果的主要条件进行优化筛选，以期建立最佳的SSR-PCR反应体系。结果表明：20 μL反应体系含有1.5 mmol/L Mg2+，60 ng DNA，0.20 mmol/L dNTPs，0.25 mmol/LPrimer和1.5 U Taq酶，最佳退火温度为44.1℃。
- Abstract:
- Taking 3 dwarf inbred line and 2 sprawling inbred lines of Cueurbita moschata as materials,the specific SSR primers in Cucurbita moschata were designed using software Primer 5.0.The main conditions affecting SSR-PCR were optimized using orthogonal experiment and gradient PCR experiments so as to establish the optimal SSR-PCR reaction system.The results showed that 20 μL reaction system contained 1.5 mmol/L Mg2+，60 ng DNA，0.20 mmol/L dNTPs，0.25 mmol/L primer and 1.5 U Taq enzyme.The optimal annealing temperature was 44.1℃.
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备注/Memo
第一作者简介：孟令君(1987-)，男，硕士，研究方向为植物生物技术。E-mail:menglingjun01987@163.com.
责任作者：李柱刚(1972)，男，博士，研究员，研究方向为植物分子生物学。
基金项目：黑龙江省杰出青年科学基金资助项目(JC200704)。收稿日期：2013-01-17