Highly sensitive and specific microRNA expression profiling using BeadArray technology.
Highly sensitive and specific microRNA expression profiling using BeadArray technology.
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DOI：
10.1093/nar/gkn387
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发表时间：
2008-08
影响因子：
14.9
通讯作者：
Fan, Jian-Bing
中科院分区：
文献类型：
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作者：
Chen, Jing;Lozach, Jean;Wickham, Eliza;Barnes, Garcia Bret;Luo, Shujun;Mikoulitch, Ivan;Zhou, Lixin;Schroth, Gary;Fan, Jian-Bing
我们开发了一种高度灵敏、特异和可重复的方法，用于微RNA（miRNA）表达谱分析，使用BeadArray™技术。该方法结合了酶辅助特异性步骤，固相引物延伸以区分miRNA家族的成员。此外，在阵列杂交之前，使用通用PCR来扩增所有靶标。目前，检测探针被设计为同时分析735种注释良好的人类miRNA。使用这种方法，用100-200 ng总RNA输入产生高度可重复的miRNA表达谱。此外，用总RNA和富集的小RNA种类获得非常相似的表达谱（R2 ≥ 0.97）。该方法具有3.5-4 log（105-109个分子）的动态范围，能够检测样品之间的1.2- 1.3倍差异。通过该方法产生的表达谱与RT-PCR（R 2 = 0.85-0.90）和直接测序（R = 0.87-0.89）获得的表达谱高度可比。该方法与96样本阵列矩阵相结合，应证明可用于大量组织样本中miRNA的高通量表达谱分析。
We have developed a highly sensitive, specific and reproducible method for microRNA (miRNA) expression profiling, using the BeadArray™ technology. This method incorporates an enzyme-assisted specificity step, a solid-phase primer extension to distinguish between members of miRNA families. In addition, a universal PCR is used to amplify all targets prior to array hybridization. Currently, assay probes are designed to simultaneously analyse 735 well-annotated human miRNAs. Using this method, highly reproducible miRNA expression profiles were generated with 100–200 ng total RNA input. Furthermore, very similar expression profiles were obtained with total RNA and enriched small RNA species (R2 ≥ 0.97). The method has a 3.5–4 log (105–109 molecules) dynamic range and is able to detect 1.2- to 1.3-fold-differences between samples. Expression profiles generated by this method are highly comparable to those obtained with RT–PCR (R2 = 0.85–0.90) and direct sequencing (R = 0.87–0.89). This method, in conjunction with the 96-sample array matrix should prove useful for high-throughput expression profiling of miRNAs in large numbers of tissue samples.
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影响因子：
64.8
作者：
Lu, J;Getz, G;Golub, TR
通讯作者：
Golub, TR
影响因子：
14.9
作者：
Chen C;Ridzon DA;Broomer AJ;Zhou Z;Lee DH;Nguyen JT;Barbisin M;Xu NL;Mahuvakar VR;Andersen MR;Lao KQ;Livak KJ;Guegler KJ
通讯作者：
Guegler KJ
影响因子：
1.7
作者：
Chudin, E.;Kruglyak, S.;McDaniel, T. K.
通讯作者：
McDaniel, T. K.
影响因子：
56.9
作者：
Kim, Jongpil;Inoue, Keiichi;Abeliovich, Asa
通讯作者：
Abeliovich, Asa
DOI：
10.1101/sqb.2003.68.69
发表时间：
2003-01-01
期刊：
COLD SPRING HARBOR SYMPOSIA ON QUANTITATIVE BIOLOGY
影响因子：
--
作者：
Fan, JB;Oliphant, A;Chee, MS
通讯作者：
Chee, MS