采用实时荧光定量逆转录聚合酶链反应(qRT-PCR)分析血浆和血清中的循环 microRNA 生物标志物。
Analysis of circulating microRNA biomarkers in plasma and serum using quantitative reverse transcription-PCR (qRT-PCR).
机构信息
Human Biology Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
出版信息
Methods. 2010 Apr;50(4):298-301. doi: 10.1016/j.ymeth.2010.01.032. Epub 2010 Feb 8.
Abstract
MicroRNAs (miRNAs) are small (approximately 22 nt) RNAs that play important roles in gene regulatory networks by binding to and repressing the activity of specific target mRNAs. Recent studies have indicated that miRNAs circulate in a stable, cell-free form in the bloodstream and that the abundance of specific miRNAs in plasma or serum can serve as biomarkers of cancer and other diseases. Measurement of circulating miRNAs as biomarkers is associated with some special challenges, including those related to pre-analytic variation and data normalization. We describe here our procedure for qRT-PCR analysis of circulating miRNAs as biomarkers, and discuss relevant issues of sample preparation, experimental design and data analysis.
摘要
微小 RNA（miRNA）是小的（大约 22 个核苷酸）RNA，通过与特定靶 mRNA 结合并抑制其活性，在基因调控网络中发挥重要作用。最近的研究表明，miRNA 以稳定的、无细胞的形式在血液中循环，血浆或血清中特定 miRNA 的丰度可以作为癌症和其他疾病的生物标志物。作为生物标志物的循环 miRNA 的测量与一些特殊的挑战有关，包括与分析前变异和数据归一化相关的挑战。我们在这里描述了我们作为生物标志物的循环 miRNA 的 qRT-PCR 分析程序，并讨论了样品制备、实验设计和数据分析的相关问题。
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