A strategy for co-analysis of microRNAs and DNA
A strategy for co-analysis of microRNAs and DNA
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microRNA 和 DNA 共同分析的策略
DOI：
10.1016/j.fsigen.2014.04.011
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发表时间：
2014-09-01
影响因子：
3.1
通讯作者：
Hou, Yiping
中科院分区：
文献类型：
--
作者：
Li, Yan;Zhang, Ji;Hou, Yiping
MicroRNA（miRNAs，长度为18-25个碱基）是一类小的非编码RNA，在转录后水平调控基因表达。小尺寸使它们比传统的mRNA更稳定，作为生物标志物识别降解或受损样品中的体液。然而，与mRNA相比，由于miRNAs的长度非常短，设计合适的引物以实现miRNAs/DNA共提取和共分析是一个挑战。在这里，我们描述了一个特定的线性RT引物的逆转录反应和一对PCR引物的设计，用于终点PCR反应的每个miRNA标记，并提出了一个策略，一组miRNA和DNA的共同分析在同一提取物使用相同的检测平台。采用毛细管电泳技术对4个miRNA标记（月经血标记miR 214、静脉血标记miR 451 a、精液标记miR 888和miR 891 a）和DNA STR复合检测试剂盒进行联合检测。结果表明，miRNAs/DNA联合分析策略不仅可以对体液进行鉴定，而且可以获得同一样本的STR图谱。(C)2014爱思唯尔爱尔兰有限公司版权所有。
MicroRNAs (miRNAs, 18-25 bases in length) are small, non-coding RNAs that regulate gene expression at the post-transcriptional level. The small size makes them more stable than conventional mRNA as a biomarker identifying body fluids in degraded or compromised samples. Compared to mRNA, however, due to the very short length of miRNAs, it is a challenge to design proper primers to achieve miRNAs/DNA co-extraction and co-analysis. Here we describe the design of a specific linear RT primer for the reverse transcription reaction and a pair of PCR primers to be used in the endpoint PCR reaction for each miRNA marker, and presented a strategy for co-analysis of a set of miRNAs and DNA on the same extract using the same detection platform. A set of 4 miRNA markers (miR214 as menstrual blood marker, miR451a as venous blood marker, miR888 and miR891a as semen markers) and a DNA STR multiplex kit were co-analyzed by capillary electrophoresis. The result demonstrated that the strategy of co-analysis of miRNAs/DNA could not only identify the body fluid, but also obtain a STR profile for the same sample. (C) 2014 Elsevier Ireland Ltd. All rights reserved.