Title | Direct Comparative Analyses of 10X Genomics Chromium and Smart-seq2 |
Authors | Wang, Xiliang He, Yao Zhang, Qiming Ren, Xianwen Zhang, Zemin |
Affiliation | Peking Univ, BIOPIC, Beijing Adv Innovat Ctr Genom, Beijing 100871, Peoples R China Peking Univ, Sch Life Sci, Beijing 100871, Peoples R China Peking Univ, Acad Adv Interdisciplinary Studies, Peking Tsinghua Ctr Life Sci, Beijing 100871, Peoples R China |
Keywords | CELL RNA-SEQ SINGLE-CELL GENE-EXPRESSION TRANSCRIPTIONAL HETEROGENEITY SPATIAL RECONSTRUCTION DESIGN POWER |
Issue Date | Apr-2021 |
Publisher | GENOMICS PROTEOMICS & BIOINFORMATICS |
Abstract | Single-cell RNA sequencing (scRNA-seq) is generally used for profiling transcriptome of individual cells. The droplet-based 10X Genomics Chromium (10X) approach and the plate-based Smart-seq2 full-length method are two frequently used scRNA-seq platforms, yet there are only a few thorough and systematic comparisons of their advantages and limitations. Here, by directly comparing the scRNA-seq data generated by these two platforms from the same samples of CD45(-) cells, we systematically evaluated their features using a wide spectrum of analyses. Smart-seq2 detected more genes in a cell, especially low abundance transcripts as well as alternatively spliced transcripts, but captured higher proportion of mitochondrial genes. The composite of Smart-seq2 data also resembled bulk RNA-seq data more. For 10X-based data, we observed higher noise for mRNAs with low expression levels. Approximately 10%-30% of all detected transcripts by both platforms were from non-coding genes, with long non-coding RNAs (lncRNAs) accounting for a higher proportion in 10X. 10X-based data displayed more severe dropout problem, especially for genes with lower expression levels. However, 10X-data can detect rare cell types given its ability to cover a large number of cells. In addition, each platform detected distinct groups of differentially expressed genes between cell clusters, indicating the different characteristics of these technologies. Our study promotes better understanding of these two platforms and offers the basis for an informed choice of these widely used technologies. |
URI | http://hdl.handle.net/20.500.11897/629501 |
ISSN | 1672-0229 |
DOI | 10.1016/j.gpb.2020.02.005 |
Indexed | SCI(E) |
Appears in Collections: | 生物医学前沿创新中心 生命科学学院 前沿交叉学科研究院 |