| 引用本文： |
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廖思明,阎冰,陈剑锋,兰国宝,文雪.对虾白斑病毒PCR反应体系的改进[J].广西科学,2004,11(2):151-153. [点击复制]
- Liao Siming,Yan Bing,Chen Jianfeng,Lan Guobao,Wen Xue.Improvement of Reaction System of PCR Applied in the Detection of White Spot Virus of Penaeid Shrimp[J].Guangxi Sciences,2004,11(2):151-153. [点击复制]
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| 摘要: |
| 采用TN匀浆,蛋白酶K、DS消化、裂解,煮沸的方法,从感染白斑病毒的养殖对虾中提取DNA作模板,在模板和扩增缓冲液用量恒定下,改变PCR反应体系的dNTP、引物和Taq酶用量,进行PCR扩增,使用琼脂糖凝胶电泳对扩增产物进行检测。结果表明,dNTP用量在125~187μM·L-1、引物用量在0.6μM·L-1、Taq酶用量达到0.5u及以上时,扩增条带荧光很强,而使用产品提供商建议用量,PCR可现特异性扩增,但不是最佳的扩增 |
| 关键词: 对虾 白斑病毒 PCR 反应体系 改进 |
| DOI： |
| 投稿时间：2003-09-24修订日期：2004-02-11 |
| 基金项目:广西科技攻关项目(桂科攻0235018-1)资助。 |
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| Improvement of Reaction System of PCR Applied in the Detection of White Spot Virus of Penaeid Shrimp |
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Liao Siming1, Yan Bing1,2, Chen Jianfeng1, Lan Guobao1, Wen Xue2
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| (1.Guangxi Mangrove Reaserch Center, 92 East Changqinlu, Beihai, Guangxi, 536000, China;2.Guangxi Institute of Oceanography, 92 East Changqinlu, Beihai, Guangxi, 536000, China) |
| Abstract: |
| Genome DNA was extracted from the tissue of the white spot virus infected cultured shrimp Penaeus monodon by TN homogenizing,DS digesting,proteinase K decomposing and boiling. Under the condition of the same amount in DNA template and the same volume in amplifying buffer,modifications were made in the reaction with the use of quantity of dNTP,primer and Taq polymerase.After PCR amplification,the products were detected by agarose electrophoresis.It shows that the fluorescent intensities of the amplified bands were much high when the quantity of dNTP was between 125 μM·L-1 and 187 μM·L-1,correspondingly the quantity of primer being 0.6 μM·L-1,and the quantity of Taq polymerase reaching 0.5 u and above.Results also showed that using the producer's recommended quantity of dNTP,primer and Taq polymerase,PCR could take the specific amplification,but not the best. |
| Key words: shrimp white spot virus PCR reaction system improvement |
