UCSD researchers have assessed the performance of seven normalization methods for single cell RNA-seq using data generated from dilution of RNA samples. Their analyses showed that methods considering spike-in ERCC RNA molecules significantly outperformed those not considering ERCCs. This work provides a guidance of selecting normalization methods to remove technical noise in single cell RNA-seq data.
Hierarchical clustering of HBR and UHR RNA-seq samples with different normalization methods not considering ERCC. (a) DESeq with HBR samples; (b) RUVr with HBR samples (c-d) DESeq and RUVr with UHR samples.