Genome biology · 2019 · 538 citations · 59 references
Life SciencesBioinformaticsBiostatisticsGene ExpressionGene Expression AnalysisTranscriptomicsCell ClusteringDimensionality ReductionSingle-cell AnalysisSingle-cell GenomicsSingle-cell Rna Sequencing AnalysisSingle-cell TranscriptomicsGround-truth ValidationHighly Variable Gene SelectionMultinomial RegressionNormalizationNormalization Of Single-cell DataRna Sequencing (Rna-seq)Single-cell Gene Expression ProfilingSingle-cell Rna Sequencing
Single-cell RNA-Seq (scRNA-Seq) profiles gene expression of individual cells. Recent scRNA-Seq datasets have incorporated unique molecular identifiers (UMIs). Using negative controls, we show UMI counts follow multinomial sampling with no zero inflation. Current normalization procedures such as log of counts per million and feature selection by highly variable genes produce false variability in dimension reduction. We propose simple multinomial methods, including generalized principal component analysis (GLM-PCA) for non-normal distributions, and feature selection using deviance. These methods outperform the current practice in a downstream clustering assessment using ground truth datasets.
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Highly Parallel Genome-wide Expression Profiling of Individual Cells Using Nanoliter Droplets
Evan Z. Macosko, Anindita Basu, Rahul Satija et al. · Cell · 2015 · 7.8K citations · Full text