适合于miRNA的荧光定量PCR优化体系的建立及其标准

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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Published in植物遗传资源学报 Vol. 13; no. 3; pp. 435 - 442
Main Author 秦剑英 刘灶长 孔德艳 周立国 罗利军
Format Journal Article
LanguageChinese
Published 上海海洋大学,上海201306%上海市农业生物基因中心,上海,201106 2012
上海市农业生物基因中心,上海201106
Subjects
MIQE标准
miRNA
基因表达量分析
荧光定量PCR
miRNA
基因表达量分析
MIQE标准
荧光定量PCR
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Summary: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。
Bibliography:Fluorescent quantitative PCR is one of the first choices for the methods in analyzing gene expression level.It is simple for 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 results indicates that the concentration of primers plays a crucial role in optimizing the fluorescence qPCR detection system.
Gene expression analysis; miRNA; Standards for MIQP; RT-qPCR
QIN Jian-ying,LIU Zao-chang,KONG De-yan,ZHOU Li-guo,LUO Li-jun（1 Shanghai Agrobiological Gene Center,Shanghai201106;2 Shanghai Ocean University,Shanghai 201306）
11-4996/S
ISSN:1672-1810
DOI:10.3969/j.issn.1672-1810.2012.03.018