| 摘要: |
| 采用CTAB-DNA提取方法,从草珊瑚植物的嫩叶中提取总DNA。以此DNA为模板,优化了草珊瑚RAPD-PCR的反应条件。结果表明,PCR扩增体系最适宜的条件为:反应体积25 μL,内含2.5 mmol/L Mg2+、1.0U DNA聚合酶、0.4 μmol/L引物、60 ng模板DNA和0.16 mmol/L dNTP。扩增程序为:94 ℃预变性2 min; 94 ℃变性30 s,37 ℃复性30 s,72 ℃延伸80 s,40个循环; 72 ℃延伸10 min; 4 ℃保存10 min。 |
| 关键词: 草珊瑚 RAPD 条件优化 |
| DOI： |
| 分类号:Q943 |
| 基金项目: |
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| Optimization of RAPD amplified conditions in medicinal plant Sarcandra glabra |
|
ZHANG Zhi-Yong1, HE Ping2,3
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1.Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Mengla 666303, China;2.College
of Life Sciences,Southwest University, Chongqing 400715, China;3.Key Laboratory of
Eco-Environment in Three-Gorges Reservior Area, Chongqing 400715, China
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| Abstract: |
| The method CTAB-DNA isolation was optimized and used to extract genomic DNA from the tender leaves of Sarcandra glabra. Based on the genomic DNA, some essential factors that affect the result of RAPD were compared and the optimal RAPD system for S.glabra was established. The results showed that the optimal concentration of five important components such as Mg2+,Taq DNA polymerase,primer,template DNA,and dNTP in 25 μL RAPD reaction system were 2.5 mmol/L,1.0U,0.4 μmol/L,60 ng,0.16 mmol/L respectively. Modified thermal profile consisted of an initial denaturation step at 94 ℃ for 2 mins,followed by 40 cycles of 94 ℃ for 30s,37 ℃ for 30 s,72 ℃ for 80 s,and a final exposure to 72 ℃ for 10mins.Then stored in 4 ℃ for 10 mins. |
| Key words: Sarcandra glabra RAPD optimization |
