Erratum: Comprehensive multi-center assessment of small RNA-seq methods for quantitative miRNA profiling
Erratum: Comprehensive multi-center assessment of small RNA-seq methods for quantitative miRNA profiling
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DOI：
10.1038/nbt0918-899b
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发表时间：
2018-10
影响因子：
46.9
通讯作者：
M. Giraldez;Ryan M Spengler;A. Etheridge;Paula M. Godoy;A. Barczak;Srimeenakshi Srinivasan;P. D. Hoff-
中科院分区：
文献类型：
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作者：
M. Giraldez;Ryan M Spengler;A. Etheridge;Paula M. Godoy;A. Barczak;Srimeenakshi Srinivasan;P. D. Hoff-
RNA-seq越来越多地用于各种样品类型（包括分离的细胞、组织和无细胞生物体液）中的小rna（例如，microrna、pirna和snorna）的定量分析。目前使用的小RNA-seq文库制备方法的准确性和可重复性尚未得到系统的测试。在这里，我们报告了由9个实验室组成的联盟获得的结果，这些实验室独立地对合成小RNA和人类血浆来源的RNA的参考样本进行了测序。我们评估了三种商用文库制备方法，使用已定义序列的适配器和六种使用退化碱基适配器的方法。鉴定了方案特异性和序列特异性偏差，包括降低小RNA-seq准确测量微小rna中腺苷-肌苷编辑能力的偏差。我们发现通过将适配器与退化基结合在一起的库准备方法可以减轻这些偏差。使用小RNA-seq的样品间MicroRNA相对定量在不同实验室和方法中是准确和可重复的。
RNA-seq is increasingly used for quantitative profiling of small RNAs (for example, microRNAs, piRNAs and snoRNAs) in diverse sample types, including isolated cells, tissues and cell-free biofluids. The accuracy and reproducibility of the currently used small RNA-seq library preparation methods have not been systematically tested. Here we report results obtained by a consortium of nine labs that independently sequenced reference, 'ground truth' samples of synthetic small RNAs and human plasma-derived RNA. We assessed three commercially available library preparation methods that use adapters of defined sequence and six methods using adapters with degenerate bases. Both protocol- and sequence-specific biases were identified, including biases that reduced the ability of small RNA-seq to accurately measure adenosine-to-inosine editing in microRNAs. We found that these biases were mitigated by library preparation methods that incorporate adapters with degenerate bases. MicroRNA relative quantification between samples using small RNA-seq was accurate and reproducible across laboratories and methods.