HCA Data Explorer

Systematic comparative analysis of single cell RNA-sequencing methods

Updated January 19, 2022

A multitude of single-cell RNA sequencing methods have been developed in recent years, with dramatic advances in scale and power, and enabling major discoveries and large scale cell mapping efforts. However, these methods have not been systematically and comprehensively benchmarked. Here, we directly compare seven methods for single cell and/or single nucleus profiling from three types of samples – cell lines, peripheral blood mononuclear cells and brain tissue – generating 36 libraries in six separate experiments in a single center. To analyze these datasets, we developed and applied scumi, a flexible computational pipeline that can be used for any scRNA-seq method. We evaluated the methods for both basic performance and for their ability to recover known biological information in the samples. Our study will help guide experiments with the methods in this study as well as serve as a benchmark for future studies and for computational algorithm development.

Joshua Z LevinThe Broad Institute of MIT & Harvardjlevin@broadinstitute.org
Jiarui Ding (Experimental Scientist)1
Xian Adiconis (Experimental Scientist)1
Sean K Simmons (Experimental Scientist)1
Monika S Kowalczyk (Experimental Scientist)1
Cynthia C Hession (Experimental Scientist)1
Nemanja D Marjanovic (Experimental Scientist)1
Travis K Hughes (Experimental Scientist)1
Marc H Wadsworth (Experimental Scientist)1
Tyler Burks (Experimental Scientist)1
Lan T Nguyen (Experimental Scientist)1
John Y Kwon (Experimental Scientist)1
Boaz Barak (Experimental Scientist)2
William Ge (Experimental Scientist)1
Amanda J Kedaigle (Experimental Scientist)1
Shaina Carroll (Experimental Scientist)1
Shuqiang Li (Experimental Scientist)1
Nir Hacohen (Experimental Scientist)1
Orit Rozenblatt-Rosen (Experimental Scientist)1
Alex K Shalek (Experimental Scientist)1
Alexandra-Chloé Villani (Experimental Scientist)1
Aviv Regev (Experimental Scientist)1
Joshua Z Levin (Principal Investigator)1
1The Broad Institute of MIT & Harvard
2MIT
Chris Villarreal
William G Sullivan
Parisa Nejad

To reference this project, please use the following link:

https://explore.data.humancellatlas.org/projects/88ec040b-8705-4f77-8f41-f81e57632f7d

Supplementary links are provided by contributors and represent items such as additional data which can’t be hosted here; code that was used to analyze this data; or tools and visualizations associated with this specific dataset.

1.https://bitbucket.org/jerry00/scumi-dev/src/master/
INSDC Project Accessions:GEO Series Accessions:

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Analysis Portals

None

Project Label

scRNAseqSystemicComparison

Species

2 species

Sample Type

specimens

Anatomical Entity

2 anatomical entities

Organ Part

2 organ parts

Selected Cell Types

mononuclear cell

Disease Status (Specimen)

normal

Disease Status (Donor)

normal

Development Stage

2 development stages

Library Construction Method

9 library construction methods

Nucleic Acid Source

2 nucleic acid sources

Paired End

false, true

Analysis Protocol

MultiSampleSmartSeq2_v2.2.6, SmartSeq2SingleSample_v5.1.5, optimus_post_processing_v1.0.0, optimus_v4.2.2

File Format

7 file formats

Cell Count Estimate

59.8k

Donor Count

4
bai2,304 file(s)bam2,310 file(s)fastq.gz6,567 file(s)loom11 file(s)mtx.gz3 file(s)tsv.gz8 file(s)txt2 file(s)