A curated list of bioinformatics benchmarking papers and resources.
The credit for this format goes to Sean Davis for his awesome-single-cell repository and Ming Tang for his ChIP-seq-analysis repository.
If you have a benchmarking study that is not yet included on this list, please make a Pull Request.
- Rules for Included Papers
- Format & Organization
- Tool/Method Sections
- Contributors
- Papers must be objective comparisons of 3 or more tools/methods.
- Papers must be awesome. This list isn't meant to chronicle every benchmarking study ever performed, only those that are particularly useful or well done.
- Papers should not be from authors showing why their tool/method is better than others.
- Benchmarking data should be publicly available or simulation code/methods must be well-documented and reproducible.
Additional guidelines/rules may be added as necessary.
Please include the following information when adding papers.
Title:
Authors:
Journal Info:
Description:
Tools/methods compared:
Recommendation(s):
Additional links (optional):
Papers within each section should be ordered by publication date, with more recent papers listed first.
Additional sections/sub-sections can be added as needed.
Title: Features that define the best ChIP-seq peak calling algorithms
Authors: Reuben Thomas, et al.
Journal Info: Briefings in Bioinformatics, May 2017
Description: This paper compared six peak calling methods on 300 simulated and three real ChIP-seq data sets across a range of significance values. Methods were scored by sensitivity, precision, and F-score.
Tools/methods compared: GEM
, MACS2
, MUSIC
, BCP
, Threshold-based method (TM)
, ZINBA
.
Recommendation(s): Varies. BCP and MACS2 performed the best across all metrics on the simulated data. For Tbx5 ChIP-seq, GEM performed the best, with BCP also scoring highly. For histone H3K36me3 and H3K4me3 data, all methods performed relatively comparably with the exception of ZINBA, which the authors could not get to run properly. MUSIC and BCP had a slight edge over the others for the histone data.
More generally, they found that methods that utilize variable window sizes and Poisson test to rank peaks are more powerful than those that use a Binomial test.
Title: A Comparison of Peak Callers Used for DNase-Seq Data
Authors: Hashem Koohy, et al.
Journal Info: PLoS ONE, May 2014
Description: This paper compares four peak callers specificity and sensitivity on DNase-seq data from two publications composed of three cell types, using ENCODE data for the same cell types as a benchmark. The authors tested multiple parameters for each caller to determine the best settings for DNase-seq data for each.
Tools/methods compared: F-seq
, Hotspot
, MACS2
, ZINBA
.
Recommendation(s): F-seq was the most sensitive, though MACS2 and Hotspot both performed competitively as well. ZINBA was the least performant by a massive margin, requiring much more time to run, and was also the least sensitive.
Title: Alignment and mapping methodology influence transcript abundance estimation
Authors: Avi Srivastava* & Laraib Malik* et al.
Journal Info: bioRXiv, October 2019
Description: This paper compares the influence of mapping and alignment on the accuracy of transcript quantification in both simulated and experimental data, as well as the effect on subsequent differential expression analysis.
Tools/methods compared: Bowtie2
, STAR
, quasi-mapping
, Selective Alignment
, RSEM
, Salmon
.
Recommendation(s): When trying to choose an approach, a choice can be made by the user performing the analysis based on any time-accuracy tradeoff they wish to make. In terms of speed, quasi-mapping is the fastest approach, followed by Selective Alignment (SA) then STAR. Bowtie2 was considerably slower than all three of these approaches. However, in terms of accuracy, SA yielded the best results, followed by alignment to the genome (with subsequent transcriptomic projection) using STAR and SA (using carefully selected decoy sequences). Bowtie2 generally performed similarly to SA, but without the benefit of decoy sequences, seemed to admit more spurious mappings. Finally, lightweight mapping of sequencing reads to the transcriptome showed the lowest overall consistency with quantifications derived from the oracle alignments. Note: Both Selective Alignment and quasi-mapping are part of salmon codebase.
Title: Statistical models for RNA-seq data derived from a two-condition 48-replicate experiment
Authors: Marek Gierliński*, Christian Cole*, Pietá Schofield*, Nicholas J. Schurch*, et al.
Journal Info: Bioinformatics, November 2015
Description: This paper compares the effect of normal, log-normal, and negative binomial distribution assumptions on RNA-seq gene read-counts from 48 RNA-seq replicates.
Tools/methods compared: normal
, log-normal
, negative binomial
.
Recommendation(s): Assuming a normal distribution leads to a large number of false positives during differential gene expression. A log-normal distribution model works well unless a sample contains zero counts. Use tools that assume a negative binomial distribution (edgeR
, DESeq
, DESeq2
, etc).
Authors: Marie-Agnès Dillies, et al.
Journal Info: Briefings in Bioinformatics, November 2013
Description: This paper compared seven RNA-seq normalization methods in the context of differential expression analysis on four real datasets and thousands of simulations.
Tools/methods compared: Total Count (TC)
, Upper Quartile (UQ)
, Median (Med),
DESeq
, edgeR
, Quantile (Q)
, RPKM
.
Recommendation(s): The authors recommend DESeq (DESeq2 now available as well) or edgeR, as those methods are robust to the presence of different library sizes and compositions, whereas the (still common) Total Count and RPKM methods are ineffective and should be abandoned.
Authors: Kimon Froussios*, Nick J Schurch*, et al.
Journal Info: Bioinformatics, February 2019
Description: This paper compared nine differential gene expression tools (and their underlying model distributions) in 17 RNA-seq replicates of Arabidopsis thaliana. Handling of inter-replicate variability and false positive fraction were the benchmarking metrics used.
Tools/methods compared: baySeq
, DEGseq
, DESeq
, DESeq2
, EBSeq
, edgeR
, limma
, Poisson-Seq
, SAM-Seq
.
Recommendation(s): Six of the tools that utilize negative binomial or log-normal distributions (edgeR, DESeq2, DESeq, baySeq, limma, and EBseq control their identification of false positives well.
Additional links: The authors released their benchmarking scripts on Github.
Authors: Nicholas J. Schurch*, Pietá Schofield*, Marek Gierliński*, Christian Cole*, Alexander Sherstnev*, et al.
Journal Info: RNA, March 2016
Description: This paper compared 11 differential expression tools on varying numbers of RNA-seq biological replicates (3-42) between two conditions. Each tool was compared against itself as a standard (using all replicates) and against the other tools.
Tools/methods compared: baySeq
, cuffdiff
, DEGSeq
, DESeq
, DESeq2
, EBSeq
, edgeR (exact and glm modes)
, limma
, NOISeq
, PoissonSeq
, SAMSeq
.
Recommendation(s): With fewer than 12 biological replicates, edgeR and DESeq2 were the top performers. As replicates increased, DESeq did a better job minimizing false positives than other tools.
Additionally, the authors recommend at least six biological replicates should be used, rising to at least 12 if users want to identify all significantly differentially expressed genes no matter the fold change magnitude.
Title: Comprehensive evaluation of differential gene expression analysis methods for RNA-seq data
Authors: Franck Rapaport, et al.
Journal Info: Genome Biology, September 2013
Description: This paper compared six differential expression methods on three cell line data sets from ENCODE (GM12878, H1-hESC, and MCF-7) and two samples from the SEQC study, which had a large fraction of differentially expressed genes validated by qRT-PCR. Specificity, sensitivity, and false positive rate were the main benchmarking metrics used.
Tools/methods compared: Cuffdiff
, edgeR
, DESeq
, PoissonSeq
, baySeq
, limma
.
Recommendation(s): Though no method emerged as favorable in all conditions, those that used negative binomial modeling (DESeq, edgeR, baySeq) generally performed best.
The more replicates, the better. Replicate numbers (both biological and technical) have a greater impact on differential detection accuracy than sequencing depth.
Title: A survey of software for genome-wide discovery of differential splicing in RNA-Seq data
Authors: Joan E Hooper
Journal Info: Human Genomics, January 2014
Description: This paper compares the methodologies, advantages, and disadvantages of eight differential splicing analysis tools, detailing use-cases and features for each.
Tools/methods compared: Cuffdiff2
, MISO
, DEXSeq
, DSGseq
, MATS
, DiffSplice
, Splicing compass
, AltAnalyze
.
Recommendation(s): This is a true breakdown of each tools' advantages and disadvantages.
The author makes no recommendation due to the performance reliance on experimental setup, data type (e.g. AltAnalyze
works best on junction + exon microarrays), and user objectives.
Table 1 provides a good comparison of the features and methodology of each method.
Authors: Gabriela A Merino
Journal Info: Briefings in Bioinformatics, March 2019
Description: This paper compares nine most commonly used workflows to detect differential isoform expression and splicing.
Tools/methods compared: EBSeq
, DESeq2
, NOISeq
, Limma
, LimmaDS
, DEXSeq
, Cufflinks
, CufflinksDS
, SplicingCompass
.
Recommendation(s): DESeq2, Limma and NOISeq for differential isoform expression(DIE) analysis and DEXSeq and LimmaDS for differential splicing (DS) testing.
Authors: Katharina E. Hayer et al.
Journal Info: Bioinformatics, Dec 2015
Description: This paper compared both guided and de novo transcript reconstruction algorithms using simulated and in vitro transcription (IVT) generated libraries. Precision/recall metrics were obtained by comparing the reconstructed transcripts to their true models.
Tools/methods compared: Cufflinks
, CLASS
, FlipFlop
, IReckon
, IsoLasso
, MiTie
, StringTie
, StringTie-SR
, AUGUSTUS
, Trinity
, SOAP
, Trans-ABySS
.
Recommendation(s): All tools measured produced less than ideal precision-recall (both <90%) when using imperfect simulated or IVT data and genes producing mulitple isoforms. Cufflinks and StringTie are among the best performers.
Title: Comprehensive evaluation of transcriptome-based cell-type quantification methods for immuno-oncology
Authors: Markus List*, Tatsiana Aneichyk*, et al.
Journal Info: Bioinformatics, July 2019
Description: This paper benchmarks and compares seven methods for computational deconvolution of cell-type abundance in bulk RNA-seq samples. Each method was tested on both simulated and true bulk RNA-seq samples validated by FACS.
Tools/methods compared: quanTIseq
, TIMER
, CIBERSORT
, CIBERSORT abs. mode
, MCPCounter
, xCell
, EPIC
.
Recommendation(s): Varies. In general, the authors recommend EPIC and quanTIseq due to their overall robustness and absolute (rather than relative) scoring, though xCell is recommended for binary presence/absence of cell types and MCPcounter was their recommended relative scoring method.
Additional links: The authors created an R package called immunedeconv for easy installation and use of all these methods. For developers, they have made available their benchmarking pipeline so that others can reproduce/extend it to test their own tools/methods.
Authors: Jiayun Chen, et al.
Journal Info: Scientific Reports, June 2019
Description: This paper compared three variant callers for WGS and WES samples from NA12878 across five next-gen sequencing platforms
Tools/methods compared: GATK
, Strelka2
, Samtools-Varscan2
.
Recommendation(s): Though all methods tested generally scored well, Strelka2 had the highest F-scores for both SNP and indel calling in addition to being the most computationally performant.
Title: Comparison of three variant callers for human whole genome sequencing
Authors: Anna Supernat, et al.
Journal Info: Scientific Reports, December 2018
Description: The paper compared three variant callers for WGS samples from NA12878 at 10x, 15x, and 30x coverage.
Tools/methods compared: DeepVariant
, GATK
, SpeedSeq
.
Recommendation(s): All methods had similar F-scores, precision, and recall for SNP calling, but DeepVariant scored higher across all metrics for indels at all coverages.
Title: A Comparison of Variant Calling Pipelines Using Genome in a Bottle as a Reference
Authors: Adam Cornish, et al.
Journal Info: BioMed Research International, October 2015
Description: This paper compared 30 variant calling pipelines composed of six different variant callers and five different aligners on NA12878 WES data from the "Genome in a Bottle" consortium.
Tools/methods compared:
- Variant callers:
FreeBayes
,GATK-HaplotypeCaller
,GATK-UnifiedGenotyper
,SAMtools mpileup
,SNPSVM
- Aligners:
bowtie2
,BWA-mem
,BWA-sampe
,CUSHAW3
,MOSAIK
,Novoalign
.
Recommendation(s): Novoalign with GATK-UnifiedGenotyper exhibited the highest sensitivity while producing few false positives.
In general, BWA-mem was the most consistent aligner, and GATK-UnifiedGenotyper
performed well across the top aligners (BWA, bowtie2, and Novoalign).
Authors: Anne Bruun Krøigård, et al.
Journal Info: PLoS ONE, March 2016
Description: This paper performed comparisons between nine somatic variant callers on five paired tumor-normal samples from breast cancer patients subjected to WES and targeted deep sequencing.
Tools/methods compared: EBCall
, Mutect
, Seurat
, Shimmer
, Indelocator
, SomaticSniper
, Strelka
, VarScan2
, Virmid
.
Recommendation(s): EBCall, Mutect, Virmid, and Strelka (now Strelka2) were most reliable for both WES and targeted deep sequencing. EBCall was superior for indel calling due to high sensitivity and robustness to changes in sequencing depths.
Title: Comparison of somatic mutation calling methods in amplicon and whole exome sequence data
Authors: Huilei Xu, et al.
Journal Info: BMC Genomics, March 2014
Description: Using the "Genome in a Bottle" gold standard variant set, this paper compared five somatic mutation calling methods on matched tumor-normal amplicon and WES data.
Tools/methods compared: GATK-UnifiedGenotyper followed by subtraction
, MuTect
, Strelka
, SomaticSniper
, VarScan2
.
Recommendation(s): MuTect and Strelka (now Strelka2) had the highest sensitivity, particularly at low frequency alleles, in addition to the highest specificity.
Title: Benchmark of tools for CNV detection from NGS panel data in a genetic diagnostics context
Authors: José Marcos Moreno-Cabrera, et al.
Journal Info: bioRxiv, November 2019.
Description: This paper compared five germline copy number variation callers against four genetic diagnostics datasets (495 samples, 231 CNVs validated by MLPA) using both default and optimized parameters. Sensitivity, specificity, positive predictive value, negative predictive value, F1 score, and various correlation coefficients were used as benchmarking metrics.
Tools/methods compared: DECoN
, CoNVaDING
, panelcn.MOPS
, ExomeDepth
, CODEX2
.
Recommendation(s): Most tools performed well, but varied based on datasets. The authors felt DECoN and panelcn.MOPS with optimized parameters were sensitive enough to be used as screening methods in genetic dianostics.
Additional links: The authors have made their benchmarking code (CNVbenchmarkeR) available, which can be run to determine optimal parameters for each algorithm for a given user's data.
Title: An evaluation of copy number variation detection tools for cancer using whole exome sequencing data
Authors: Fatima Zare, et al.
Journal Info: BMC Bioinformatics, May 2017
Description: This paper compared six copy number variation callers on ten TCGA breast cancer tumor-normal pair WES datasets in addition to simulated datasets from VarSimLab. Sensitivity, specificity, and false-discovery rate were used as the benchmarking metrics.
Tools/methods compared: ADTEx
, CONTRA
, cn.MOPS
, ExomeCNV
, VarScan2
, CoNVEX
.
Recommendation(s): All tools suffered from high FDRs (~30-60%), but [ExomeCNV]https://github.com/cran/ExomeCNV) (a now defunct R package) had the highest overall sensitivity. VarScan2 had moderate sensitivity and specificity for both amplifications and deletions.
Authors: Daniel L. Cameron, et al.
Journal Info: Nature Communications, July 2019
Description: This paper compared 10 structural variant callers on four cell line WGS datasets (NA12878, HG002, CHM1, and CHM13) with orthogonal validation data. Precision and recall were the benchmarking metrics used.
Tools/methods compared: BreakDancer
, cortex
, CREST
, DELLY
, GRIDSS
, Hydra
, LUMPY
, manta
, Pindel
, SOCRATES
.
Recommendation(s): The authors found GRIDSS and manta consistently performed well, but also provide more general guidelines for both users and developers.
- Use a caller that utilizes multiple sources of evidence and assembly.
- Use a caller that can call all events you care about.
- Ensemble calling is not a cure-all and generally don't outperform the best individual callers (at least on these datasets).
- Do not use callers that rely only on paired-end data.
- Calls with high read counts are typically artefacts.
- Simulations aren't real - benchmarking solely on simulations is a bad idea.
- Developers - be wary of incomplete trust sets and the potential for overfitting. Test tools on multiple datasets.
- Developers - make your tool easy to use with basic sanity checks to protect against invalid inputs. Use standard file formats.
- Developers - use all available evidence and produce meaningful quality scores.
Title: Comprehensive evaluation of structural variation detection algorithms for whole genome sequencing
Authors: Shunichi Kosugi, et al.
Journal Info: Genome Biology, June 2019
Description: This study compared 69 structural variation callers on simulated and real (NA12878, HG002, and HG00514) datasets. F-scores, precision, and recall were the main benchmarking metrics.
Tools/methods compared: 1-2-3-SV
, AS-GENESENG
, BASIL-ANISE,
BatVI
, BICseq2
, BreakDancer
, BreakSeek
, BreakSeq2
, Breakway
, CLEVER
, CNVnator
,
Control-FREEC
, CREST
, DELLY
, DINUMT
, ERDS
, FermiKit
, forestSV
, GASVPro
, GenomeSTRiP
, GRIDSS
, HGT-ID
, Hydra-sv
, iCopyDAV
, inGAP-sv
, ITIS
,
laSV
, Lumpy
, Manta
, MATCHCLIP
, Meerkat
, MELT
, MELT-numt
, MetaSV
, MindTheGap
, Mobster
, Mobster-numt
, Mobster-vei
, OncoSNP-SEQ
, Pamir
, PBHoney
,
PBHoney-NGM
, pbsv
, PennCNV-Seq
, Pindel
, PopIns
, PRISM
, RAPTR
, readDepth
, RetroSeq
, Sniffles
, Socrates
, SoftSearch
, SoftSV
, SoloDel
, Sprites
,
SvABA
, SVDetect
, Svelter
, SVfinder
, SVseq2
, Tangram
, Tangram-numt
, Tangram-vei
, Tea
, TEMP
, TIDDIT
, Ulysses
, VariationHunter
, VirusFinder
, VirusSeq
, Wham
.
Recommendation(s): Varies greatly depending on type and size of the structural variant in addition to read length.
GRIDSS
, Lumpy
, SVseq2
, SoftSV
, and Manta
performed well calling deletions of diverse sizes.
TIDDIT
, forestSV
, ERDS
, and CNVnator
called large deletions well, while pbsv
, Sniffles
, and PBHoney
were the best performers for small deletions.
For duplications, good choices included Wham
, SoftSV
, MATCHCLIP
, and GRIDSS
, while CNVnator
, ERDS
, and iCopyDAV
excelled calling large duplications.
For insertions, MELT
, Mobster
, inGAP-sv
, and methods using long read data were most effective.
Title: Evaluating nanopore sequencing data processing pipelines for structural variation identification
Authors: Anbo Zhou, et al.
Journal Info: Genome Biology, November 2019
Description: This paper evaluated four alignment tools and three SV detection tools on four nanopore datasets (both simulated and real).
Tools/methods compared: aligners - minimap2
, NGMLR
, GraphMap
, LAST
. SV Callers - Sniffles
, NanoSV
, Picky
.
Recommendation(s): The authors recommend using the minimap2 aligner in combination with the SV caller Sniffles because of their speed and relatively balanced performance.
Additional links (optional): The authors provide all code used in the study as well as a singularity package containing pre-installed programs and all seven pipeline.
Title: A comparison of single-cell trajectory inference methods
Authors: Wouter Saelens*, Robrecht Cannoodt*, et al.
Journal Info: Nat Biotech, April 2019
Description: A comprehensive evaluation of 45 trajectory inference methods, this paper provides an unmatched comparison of the rapidly evolving field of single-cell trajectory inference. Each method was scored on accuracy, scalability, stability, and usability. Should be considered a gold-standard for other benchmarking studies.
Tools/methods compared: PAGA
, RaceID/StemID
, SLICER
, Slingshot
, PAGA Tree
, MST
, pCreode
, SCUBA
, Monocle DDRTree
, Monocle ICA
, cellTree maptpx
, SLICE
, cellTree VEM
, EIPiGraph
, Sincell
, URD
, CellTrails
, Mpath
, CellRouter
, STEMNET
, FateID
, MFA
, GPfates
, DPT
, Wishbone
, SCORPIUS
, Component 1
, Embeddr
, MATCHER
, TSCAN
, Wanderlust
, PhenoPath
, topslam
, Waterfall
, EIPiGraph linear
, ouijaflow
, FORKS
, Angle
, EIPiGraph cycle
, reCAT
.
Recommendation(s): Varies depending on dataset and expected trajectory type, though PAGA, PAGA Tree, SCORPIUS, and Slingshot all scored highly across all metrics.
Authors wrote an interactive Shiny app to help users choose the best methods for their data.
Additional links: The dynverse site contains numerous packages for users to run and compare results from different trajectory methods on their own data without installing each individually by using Docker. Additionally, they provide several tools for developers to wrap and benchmark their own method against those included in the study.
Title: A benchmark of batch-effect correction methods for single-cell RNA sequencing data
Authors: Hoa Thi Nhu Tran et al.
Journal Info: Genome Biology January 2020
Description: Authors compared 14 methods in terms of computational runtime, the ability to handle large datasets, and batch-effect correction efficacy while preserving cell type purity.
Tools/methods compared:
Seurat2
, Seurat3
, Harmony
, fastMNN
, MNN Correct
, ComBat
, Limma
, scGen
, Scanorama
, MMD-ResNet
, ZINB-WaVe
, scMerge
, LIGER
, BBKNN
Recommendation(s): Based on the benchmarking results authors suggest Harmony, LIGER, and Seurat3 as best methods for batch integration.
Authors: Shiquan Sun, Jiaqiang Zhu, Ying Ma, Xiang Zhou
Journal Info: BioRxiv, October 2019
Description: A mammoth comparison of 18 different dimension reduction methods on 30 publicly available scRNAseq data sets in addition to 2 simulated datasets for a variety of purposes ranging from cell clustering to trajectory inference to neighborhood preservation.
Tools/methods compared:
factor analysis (FA)
, principal component analysis (PCA)
, independent component analysis (ICA)
, Diffusion Map
, nonnegative matrix factorization (NMF)
, Poisson NMF
, zero-inflated factor analysis (ZIFA)
, zero-inflated negative binomial based wanted variation extraction (ZINB-WaVE)
, probabilistic count matrix factorization (pCMF)
, deep count autoencoder network (DCA)
, scScope
, generalized linear model principal component analysis (GLMPCA)
, multidimensional scaling (MDS)
, locally linear embedding (LLE)
, local tangent space alignment (LTSA)
, Isomap
, uniform manifold approximation and projection (UMAP)
, t-distributed stochastic neighbor embedding (tSNE)
.
Recommendation(s): Varies depending on use case. Factor Analysis and principal component analysis performed well for most use cases. See figure 5 for pratical guidelines.
Additional links: The authors have made their benchmarking code available on Github.
Authors: Fenglin Liu, Yuanyuan Zhang, et al.
Journal Info: Genome Biology, November 2019
Description: This paper compared seven variant callers using both simulation and real scRNA-seq datasets and identified several elements influencing their performance, including read depth, variant allele frequency, and specific genomic contexts. Sensitivity and specificity were the benchmarking metrics used.
Tools/methods compared: SAMtools
, GATK
, CTAT
, FreeBayes
, MuTect2
, Strelka2
, VarScan2
.
Recommendation(s): Varies, see figure 7 for a flowchart breakdown. Generally, SAMtools (most sensitive, lower specificity in intronic or high-identity regions), Strelka2 (good performance when read depth >5), FreeBayes (good specificity/sensitivity in cases with high variant allele frequencies), and CTAT (no alignment step necessary) were top performers.
Additional links: The authors made their benchmarking code available on Github.
Title: Assessment of computational methods for the analysis of single-cell ATAC-seq data
Authors: Caleb Lareau*, Tommaso Andreani*, Micheal E. Vinyard*, et al.
Journal Info: Genome Biology, November 2019
Description: This study compares 10 methods for scATAC-seq processing and featurizing using 13 synthetic and real datasets from diverse tissues and organisms.
Tools/methods compared: BROCKMAN
, chromVAR
, cisTopic
, Cicero
, Gene Scoring
, Cusanovich2018
, scABC
, Scasat
, SCRAT
, SnapATAC
.
Recommendation(s): SnapATAC, Cusanovich2018, and cisTopic were the top performers for separating cell populations of different coverages and noise levels. SnapATAC was the only method capable of analyzing a large dataset (>80k cells).
Additional links: The authors have made their benchmarking code available on Github.
Title: A practical guide to methods controlling false discoveries in computational biology
Authors: Keegan Korthauer*, Patrick K. Kimes*, et al.
Journal Info: Genome Biology, June 2019
Description: An benchmark comparison of the accuracy, applicability, and ease of use of two classic and six modern methods that control for the false discovery rate (FDR). Used simulation studies as well as six case studies in computational biology (specifically differential expression testing in bulk RNA-seq, differential expression testing in single-cell RNA-seq, differential abundance testing and correlation analysis in 16S microbiome data, differential binding testing in ChIP-seq, genome-wide association testing, and gene set analysis).
Tools/methods compared: Benjamini-Hochberg, Storey’s q-value, conditional local FDR (LFDR), FDR regression (FDRreg), independent hypothesis weighting (IHW), adaptive shrinkage (ASH), Boca and Leek’s FDR regression (BL), and adaptive p-value thresholding (AdaPT).
Recommendation(s): Modern FDR methods that use an informative covariate (as opposed to only p-values) leads to more power while controlling the FDR over classic methods. The improvement of the modern FDR methods over the classic methods increases with the informativeness of the covariate, total number of hypothesis tests, and proportion of truly non-null hypotheses.
Additional links: Full analyses of the in silico experiments, simulations, and case studies are provided in Additional files 2–41 at https://pkimes.github.io/benchmark-fdr-html/. The source code to reproduce all results in the manuscript and additional files, as well as all figures, is available on GitHub. An ExperimentHub
package containing the full set of results objects is available through the Bioconductor project, and a Shiny application for interactive exploration of these results is also available on GitHub. The source code, ExperimentHub package, and Shiny application are all made available under the MIT license.
- Jared Andrews (@j-andrews7)
- Kevin Blighe (@kevinblighe, biostars)
- Jeremy Leipzig (@leipzig)
- Avi Srivastava (@k3yavi)
- Stephanie Hicks (@stephaniehicks)
- Sridhar N Srivatsan (@sridhar0605)