| 适合于miRNA的荧光定量PCR优化体系的建立及其标准 |
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| 引用本文： | 刘灶长,秦剑英,罗利军. 适合于miRNA的荧光定量PCR优化体系的建立及其标准[J]. 植物遗传资源学报, 2012, 13(3): 435-442 |
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| 作者姓名： | 刘灶长 秦剑英 罗利军 |
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| 作者单位： | 1. 上海市农业生物基因中心,上海201106;上海海洋大学,上海201306
2. 上海市农业生物基因中心,上海,201106 |
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| 基金项目： | 国家自然科学基金项目（面上项目, No.31071400） |
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| 摘 要： | 实时荧光定量PCR是目前基因表达量差异分析的首选方法之一。虽然其操作简单,但如何保证其定量结果的可信度一直是个难题,特别是针对只有20多个碱基的miRNA的定量分析。本研究以水稻miR408在不同组织中的表达差异为实例,系统地优化和阐述了有关荧光定量的新标准及要求。结果表明,引物的浓度对于荧光定量PCR体系的优化至关重要。
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| 关 键 词： | 基因表达量分析 MIQE标准 miRNA 荧光定量PCR |
| 收稿时间： | 2011-04-01 |
| 修稿时间： | 2011-12-07 |
Establishment and Criteria of Optimal Reaction System of RT-qPCR Suitable for miRNA |
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| QIN Jian-ying , LIU Zao-chang , KONG De-yan , ZHOU Li-guo , LUO Li-jun. Establishment and Criteria of Optimal Reaction System of RT-qPCR Suitable for miRNA[J]. Journal of Plant Genetic Resources, 2012, 13(3): 435-442 |
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| Authors: | QIN Jian-ying LIU Zao-chang KONG De-yan ZHOU Li-guo LUO Li-jun |
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| Affiliation: | Shanghai Agrobiological Gene Center |
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| Abstract: | Fluorescent quantitative PCR is one of the first choices for methods in analyzing gene expression level. It is a simple way of operation, but how to ensure the credibility of the quantitative PCR results is still a problem to be tackled especially for the quantitative analysis of miRNA with only about 20 nucleotides. In this paper, the expression difference of miR408 gene in different tissues in rice was taken for instance. The new criteria and requirements of fluorescent quantitative PCR (RT-qPCR) were systematically optimized and summarized. The experimental results showed that the concentration of primers plays a crucial role in optimizing the fluorescence qPCR detection system. |
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| Keywords: | Gene Expression Analysis miRNA Standards for MIQE RT-qPCR |
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