Simple and sensitive microRNA labeling by terminal deoxynucleotidyl transferase

	Simple and sensitive microRNA labeling by terminal deoxynucleotidyl transferase
	Zhao, BT; Gong, ZZ; Ma, ZL; Wang, DT; Jin, YX
刊名	ACTA BIOCHIMICA ET BIOPHYSICA SINICA
	2012
卷号	44 期号:2 页码:129-135
关键词	microRNA microarray microRNA labeling terminal deoxynucleotidyl transferase (TdT)
通讯作者	Zhao, BT (reprint author), Shanghai Univ, Sch Life Sci, Shanghai 200444, Peoples R China.,botaozhao@gmail.com ; yxjin@sibs.ac.cn
英文摘要	MicroRNAs (miRNAs) constitute a critically important class of non-translated, small RNAs, which post-transcriptionally regulate gene expression via one of the multiple mechanisms. To profile miRNA expression, microarrays have been extensively applied to the high-throughput detection of miRNAs. Here, we described a novel 3'-end miRNA-labeling method for microarray detection by terminal-deoxynucleotidyl transferase (TdT). TdT can catalyze the formation of polynucleotides at RNA receptor molecule with deoxycytidine triphosphate (dCTP). Using this activity, miRNA was successfully labeled by adding fluorescence dCTPs to its 3'-end. This labeling method was very simple and sensitive. The TdT-labeling method can detect as little as 0.04 fmol of synthetic small RNA, and produce precise and accurate measurements that span a linear dynamic range from 0.04 to 5 fmol of synthetic small RNA. The high consistency of miRNA expression data between our TdT method and real-time polymerase chain reaction analysis indicated the reliability and accuracy of the TdT method. Taken together, these results emphasize the immense potential application of the TdT-labeling method for sensitive and high-throughput microarray analysis of miRNA expression.
学科主题	Biochemistry & Molecular Biology; Biophysics
类目[WOS]	Biochemistry & Molecular Biology ; Biophysics
关键词[WOS]	OLIGONUCLEOTIDE MICROARRAY ; GENE-EXPRESSION ; RNA ; REGULATORS ; PLATFORM ; REVEALS ; TISSUES ; MIRNAS ; FATE
收录类别	SCI
语种	英语
WOS记录号	WOS:000299744000004
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/551]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Zhao, BT,Gong, ZZ,Ma, ZL,et al. Simple and sensitive microRNA labeling by terminal deoxynucleotidyl transferase[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2012,44(2):129-135.
APA	Zhao, BT,Gong, ZZ,Ma, ZL,Wang, DT,&Jin, YX.(2012).Simple and sensitive microRNA labeling by terminal deoxynucleotidyl transferase.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,44(2),129-135.
MLA	Zhao, BT,et al."Simple and sensitive microRNA labeling by terminal deoxynucleotidyl transferase".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 44.2(2012):129-135.