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Label-free microRNA profiling not biased by 3' end 2'-O-methylation

Accurate quantification of miRNA expression level is essential to the study of its biology, and many cutting-edge technologies have been developed to accommodate this need. Yet most of them were designed primarily for the "regular" RNAs such as animal miRNAs and may overlook the fact that plant miRN... Full description

Journal Title: Analytical chemistry 07 August 2012, Vol.84(15), pp.6361-5
Main Author: Shen, Ye
Other Authors: Zheng, Ke-Xiao , Duan, Demin , Jiang, Li , Li, Jiong
Format: Electronic Article Electronic Article
Language: English
Subjects:
ID: E-ISSN: 1520-6882 ; PMID: 22779405 Version:1 ; DOI: 10.1021/ac301360z
Link: http://pubmed.gov/22779405
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recordid: medline22779405
title: Label-free microRNA profiling not biased by 3' end 2'-O-methylation
format: Article
creator:
  • Shen, Ye
  • Zheng, Ke-Xiao
  • Duan, Demin
  • Jiang, Li
  • Li, Jiong
subjects:
  • Oligonucleotide Array Sequence Analysis
  • Micrornas -- Analysis
ispartof: Analytical chemistry, 07 August 2012, Vol.84(15), pp.6361-5
description: Accurate quantification of miRNA expression level is essential to the study of its biology, and many cutting-edge technologies have been developed to accommodate this need. Yet most of them were designed primarily for the "regular" RNAs such as animal miRNAs and may overlook the fact that plant miRNAs and many other small noncoding RNAs are 2'-O-methylated at the 3' end nucleotide. According to our experimental data and previous reports, this structural variation is detrimental to the effectiveness of the commonly used enzymatic labeling methods, leading to strongly biased results (~24-fold difference). Herein, we demonstrate that our Stacking-Hybridized Universal Tag (SHUT) microarray assay is well suited for unbiased profiling of both normal and methylated small RNA species. The detected signals of small RNAs with 2'-hydroxyl and 2'-O-methyl 3' ends are highly consistent (no significant difference at α = 0.01 level). For specificity, the presented method edges over others by its unique...
language: eng
source:
identifier: E-ISSN: 1520-6882 ; PMID: 22779405 Version:1 ; DOI: 10.1021/ac301360z
fulltext: fulltext
issn:
  • 15206882
  • 1520-6882
url: Link


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titleLabel-free microRNA profiling not biased by 3' end 2'-O-methylation
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descriptionAccurate quantification of miRNA expression level is essential to the study of its biology, and many cutting-edge technologies have been developed to accommodate this need. Yet most of them were designed primarily for the "regular" RNAs such as animal miRNAs and may overlook the fact that plant miRNAs and many other small noncoding RNAs are 2'-O-methylated at the 3' end nucleotide. According to our experimental data and previous reports, this structural variation is detrimental to the effectiveness of the commonly used enzymatic labeling methods, leading to strongly biased results (~24-fold difference). Herein, we demonstrate that our Stacking-Hybridized Universal Tag (SHUT) microarray assay is well suited for unbiased profiling of both normal and methylated small RNA species. The detected signals of small RNAs with 2'-hydroxyl and 2'-O-methyl 3' ends are highly consistent (no significant difference at α = 0.01 level). For specificity, the presented method edges over others by its unique...
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abstractAccurate quantification of miRNA expression level is essential to the study of its biology, and many cutting-edge technologies have been developed to accommodate this need. Yet most of them were designed primarily for the "regular" RNAs such as animal miRNAs and may overlook the fact that plant miRNAs and many other small noncoding RNAs are 2'-O-methylated at the 3' end nucleotide. According to our experimental data and previous reports, this structural variation is detrimental to the effectiveness of the commonly used enzymatic labeling methods, leading to strongly biased results (~24-fold difference). Herein, we demonstrate that our Stacking-Hybridized Universal Tag (SHUT) microarray assay is well suited for unbiased profiling of both normal and methylated small RNA species. The detected signals of small RNAs with 2'-hydroxyl and 2'-O-methyl 3' ends are highly consistent (no significant difference at α = 0.01 level). For specificity, the presented method edges over others by its unique...
doi10.1021/ac301360z
pmid22779405
date2012-08-07