Direct multiplexed measurement of gene expression with color-coded probe pairs
Direct multiplexed measurement of gene expression with color-coded probe pairs
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DOI：
10.1038/nbt1385
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发表时间：
2008-03-01
影响因子：
46.9
通讯作者：
Hood, Leroy
中科院分区：
文献类型：
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作者：
Geiss, Gary K.;Bumgarner, Roger E.;Hood, Leroy
我们描述了一种技术，即 NanoString nCounter 基因表达系统，它可以捕获并计算单个 mRNA 转录本。相对于现有平台的优势包括直接测量 mRNA 表达水平而无需酶促反应或偏差、灵敏度与高多重能力相结合以及数字读数。对 509 个人类基因进行的实验产生了 0.999 的重复相关系数、0.1 fM 至 0.5 fM 之间的检测限以及超过 500 倍的线性动态范围。 NanoString nCounter基因表达系统与微阵列和TaqMan PCR的比较表明，nCounter系统比微阵列更灵敏，并且与实时PCR的灵敏度相似。最后，通过 nCounter 系统和 SYBR Green 实时 PCR 测量的 7 个样本中 21 个基因的转录水平进行比较，结果表明所有转录水平的基因表达模式相似。
We describe a technology, the NanoString nCounter gene expression system, which captures and counts individual mRNA transcripts. Advantages over existing platforms include direct measurement of mRNA expression levels without enzymatic reactions or bias, sensitivity coupled with high multiplex capability, and digital readout. Experiments performed on 509 human genes yielded a replicate correlation coefficient of 0.999, a detection limit between 0.1 fM and 0.5 fM, and a linear dynamic range of over 500-fold. Comparison of the NanoString nCounter gene expression system with microarrays and TaqMan PCR demonstrated that the nCounter system is more sensitive than microarrays and similar in sensitivity to real-time PCR. Finally, a comparison of transcript levels for 21 genes across seven samples measured by the nCounter system and SYBR Green real-time PCR demonstrated similar patterns of gene expression at all transcript levels.