BUTTERFLY: addressing the pooled amplification paradox with unique molecular identifiers in single-cell RNA-seq.

BUTTERFLY: addressing the pooled amplification paradox with unique molecular identifiers in single-cell RNA-seq.
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DOI:
10.1186/s13059-021-02386-z
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发表时间:
2021-06-08
期刊:
影响因子:
12.3
通讯作者:
Pachter L
Pachter L
中科院分区:
生物学1区
文献类型:
--
作者:
Gustafsson J;Robinson J;Nielsen J;Pachter L

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在单细胞RNA-seq测定中掺入独特的分子标识符(UMI)使得鉴定重复的分子成为可能,从而促进从测序读数中计数不同的分子。然而,我们发现,天真的重复删除可能会导致偏见,由于“合并扩增悖论”,我们提出了一种改进的量化方法的基础上看不见的物种建模。我们的校正称为蝴蝶使用零截断负二项估计在kallisto bustools工作流程中实现。我们证明了它在细胞类型和基因中的功效,并表明在某些情况下它可以逆转基因的相对丰度。在线版本包含补充材料,可通过10.1186/s13059-021-02386-z获得。
The incorporation of unique molecular identifiers (UMIs) in single-cell RNA-seq assays makes possible the identification of duplicated molecules, thereby facilitating the counting of distinct molecules from sequenced reads. However, we show that the naïve removal of duplicates can lead to a bias due to a “pooled amplification paradox,” and we propose an improved quantification method based on unseen species modeling. Our correction called BUTTERFLY uses a zero truncated negative binomial estimator implemented in the kallisto bustools workflow. We demonstrate its efficacy across cell types and genes and show that in some cases it can invert the relative abundance of genes. The online version contains supplementary material available at 10.1186/s13059-021-02386-z.
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