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| 运用荧光定量PCR技术检测中肋骨条藻的研究 | |
| 引用本文： | 何闪英,程刚,苟万里,李荣秀,米铁柱,于志刚.运用荧光定量PCR技术检测中肋骨条藻的研究[J].高技术通讯,2006,16(3):313-318. |
| 作者姓名： | 何闪英 程刚 苟万里 李荣秀 米铁柱 于志刚 |
| 作者单位： | 1. 中国海洋大学,青岛,266003 2. 青岛出入境检验检疫局,青岛,266101 3. 上海交通大学生命科学技术学院,上海,200030 |
| 基金项目： | 国家科技攻关项目 , 中国科学院资助项目 |
| 摘 要： | 以rDNA序列为寻找种特异性引物的靶区域,通过分析中肋骨条藻rDNA序列,设计出7套适合用于实时荧光定量PCR方法(RFQ-PCR)的引物与探针,经引物验证,选择Primer6(F/R)进一步分析,对比扩增序列及核酸杂交验证,表明该探针可作为中肋骨条藻特异性探针.最后以Primer6(F/R)和TaqMan6建立了定量检测中肋骨条藻的RFQ-PCR方法,并绘制出了定量检测中肋骨条藻的标准曲线,测定数据用镜检计数进行了验证,证明该方法是准确可靠的. |
| 关 键 词： | 中肋骨条藻 定量PCR 分子探针 |
| 收稿时间： | 2005-04-29 |
| 修稿时间： | 2005年4月29日 |
Development of a real-time PCR for rapid detection of Skeletonema costatum |
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| He Shanying,Chen Gang,Gou Wanli,Li Rongxiu,Mi Tiezhu,Yu Zhigang.Development of a real-time PCR for rapid detection of Skeletonema costatum[J].High Technology Letters,2006,16(3):313-318. | |
| Authors: | He Shanying Chen Gang Gou Wanli Li Rongxiu Mi Tiezhu Yu Zhigang |
| Abstract: | Sequences of the rDNA and ITS deduced from Skeletonema costatum was used for designing species-specific primers, and a real-time fluorescent quantitative polymerase chain reaction (RFQ-PCR) method was developed for quantitative detection of S. costatum. Seven sets of primers and probes were designed, and were applied to specificity check with PCR and dot-blot analysis. Probe 6 and primer 6 (F/R) were selected to develop a RFQ-PCR method for S. costatum. A calibration curve was constructed with Cycle Threshold value against visual counted cell number. The value of the curve was tested with other S.costatum samples, which were assayed with both RFQ-PCR method and visual count under microscope. |
| Keywords: | Skeletonema costatum quantitative PCR molecular probe |
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