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Efficient phasing and imputation of low-coverage sequencing data using large reference panels

A Publisher Correction to this article was published on 20 January 2021

This article has been updated


Low-coverage whole-genome sequencing followed by imputation has been proposed as a cost-effective genotyping approach for disease and population genetics studies. However, its competitiveness against SNP arrays is undermined because current imputation methods are computationally expensive and unable to leverage large reference panels. Here, we describe a method, GLIMPSE, for phasing and imputation of low-coverage sequencing datasets from modern reference panels. We demonstrate its remarkable performance across different coverages and human populations. GLIMPSE achieves imputation of a genome for less than US$1 in computational cost, considerably outperforming other methods and improving imputation accuracy over the full allele frequency range. As a proof of concept, we show that 1× coverage enables effective gene expression association studies and outperforms dense SNP arrays in rare variant burden tests. Overall, this study illustrates the promising potential of low-coverage imputation and suggests a paradigm shift in the design of future genomic studies.

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Fig. 1: Overview of GLIMPSE.
Fig. 2: Performance and running time of low-coverage sequencing phasing and imputation.
Fig. 3: Comparison of low-coverage and SNP array imputation.
Fig. 4: Functional variant analysis across low-coverage and SNP array call sets.

Data availability

The 1000 Genomes Project phase 3 dataset sequenced at high coverage by the New York Genome Center is available on the European Nucleotide Archive under accession no. PRJEB31736. The publicly available subset of the HRC dataset is available from the European Genome-phenome Archive at the European Bioinformatics Institute (EBI) under accession no. EGAS00001001710. The Genome in A Bottle data for sample NA12878 is available at the National Center for Biotechnology Information ftp website: The subset of the 1000 Genomes samples genotyped on Affymetrix6.0 is available at GnomAD v.3 is available at The list of positions used to simulate the SNP arrays is available at The RNA-seq data are part of the Geuvadis study and are available at the EBI ArrayExpress under accession code no. E-GEUV-1. The ENCODE project was accessed using accession nos. integration_data_jan2011 for the lymphoblastoid cell line-specific protein binding sites, ENCSR000EJD for the DNase-hypersensitive sites and ENCSR000AKC for locations with H3K27ac histone modifications. The results shown in Fig. 3a,b are a subset of the configurations tested. A full view of the results in available at the GLIMPSE website (European population:, African-American population: The full raw data used to generate Fig. 3a,b and the benchmark shown on the website are available at the GLIMPSE repository ( Source data are provided with this paper.

Code availability

GLIMPSE is available from and

Change history

  • 20 January 2021

    An amendment to this paper has been published and can be accessed via a link at the top of the paper.


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This work was funded by a Swiss National Science Foundation project grant no. PP00P3_176977. The New York Genome Center 1000 Genomes data were generated at the New York Genome Center with funds provided by a National Human Genome Research Institute grant no. 3UM1HG008901–03S1. We thank S. Carmi for useful comments on the preprint version of the manuscript.

Author information




S.R., D.M.R. and O.D. designed the study, performed the experiments and drafted the paper. S.R. and O.D. developed the algorithm and wrote the software. S.R., R.J.H. and O.D. created the website. O.D. supervised the project. All authors reviewed the final manuscript.

Corresponding author

Correspondence to Olivier Delaneau.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Peer review information Nature Genetics thanks Garrett Hellenthal, Sam Morris and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Extended Data Fig. 1 Read count distribution of downsampled sequencing data.

The y-axis shows the fractions of genotypes covered by 0 to 11 sequencing reads across multiple downsampled coverages from 0.1x to 4.0x. The color bars show the observed fractions in the downsampled data while the black dots and lines show the expected fractions assuming coverage is Poisson distributed.

Extended Data Fig. 2 Phasing performance of subsets of EUR and ASW samples.

Performance of the GLIMPSE (blue line) and SHAPEIT4 (black line) phasing algorithms. SHAPEIT4 has been run to rephase the genotype calls produced by GLIMPSE as it can only handle hard called genotypes. Validation genotypes were generated using an Affymetrix 6.0 SNP array. Validation haplotypes were derived thanks to additional samples being genotyped allowing to form multiple duos and trios.

Extended Data Fig. 3 Genotype discordance.

Genotype discordance stratified by minor-allele-frequency for the 1x coverage European population dataset on chromosome 1. The reference panel used are a subset of 5,000 samples (solid lines), and 20,000 samples from the HRC (dashed lines). The genotype discordance is shown for (A) all genotypes, and split between (B) major/major, (C) major/minor or (D) minor/minor genotypes in the validation dataset.

Extended Data Fig. 4 Zoomed-in genotype discordance for MAF > 1%.

Genotype discordance stratified by minor-allele-frequency (MAF > 1%) for the 1x coverage European population dataset on chromosome 1. The reference panels used are a subset of 5,000 samples (solid lines), and 20,000 samples from the HRC (dashed lines). The genotype discordance is shown for (A) all genotypes, and split between (B) major/major, (C) major/minor or (D) minor/minor genotypes in the validation dataset.

Extended Data Fig. 5 Non-reference discordance.

Non-reference discordance (NRD) stratified by non-reference allele frequency for the 1x coverage European population dataset on chromosome 1. The reference panels used are a subset of 5,000 samples (solid lines), and 20,000 samples from the HRC (dashed lines). (A.) Non-reference allele frequency > 0.01%; (B.) Non-reference allele frequency > 1%. The NRD is calculated as \(\left( {e_{rr} + e_{ra} + e_{aa}} \right)/\left( {m_{ra} + m_{aa} + e_{rr} + e_{ra} + e_{aa}} \right)\), where err, era and eaa are the counts of the mismatches for the homozygous reference, heterozygous and homozygous alternative genotypes, while mra and maa are the counts of the matches at heterozygous and homozygous alternative genotypes.

Extended Data Fig. 6 Calibration of genotype posteriors for 1.0x coverage.

(A.) Calibration of genotype posterior probabilities of different imputation methods for 1.0x coverage European dataset on chromosome 1. The reference panels used are a subset of 5,000 samples (solid lines), and 20,000 samples from the HRC (dashed lines). Imputed genotypes are binned according to the posterior probability distribution (x-axis) and plotted against the percentage of concordance against high coverage data (y-axis). (B.) Number of genotypes per probability bin.

Extended Data Fig. 7 Running time of imputation methods.

Running time of low-coverage sequencing imputation methods for the European population chromosome 1 dataset. We only ran GENEIMP on 1x coverage data. For BEAGLE and GENEIMP we only show reference panel size up to 5,000 samples due to time limits. The vertical axis is on a log scale.

Extended Data Fig. 8 Memory usage of imputation methods.

Memory usage of low-coverage sequencing methods for the European population chromosome 1 dataset. We only ran GENEIMP on 1x coverage data. For BEAGLE and GENEIMP we only show reference panel size up to 5,000 samples due to time limits. LOIMPUTE imputes a single sample at the time, therefore the reported memory usage is for a single sample, while we report the memory usage for the full cohort of 503 individuals for all other methods. The vertical axis is on a log scale.

Extended Data Fig. 9 Lead eQTL overlap and association p-value mean absolute error.

(A) Overlap between lead eQTLs identified in high-coverage and each low-coverage and SNP array dataset. eQTL mapping was performed independently for each dataset (FDR 5%; MAF > = 1%). eGenes in which the lead eQTL p-value was tied with another variant’s p-value (for example due to perfect linkage disequilibrium) were excluded, as the choice of variant for being the lead eQTL in these cases is arbitrary. The total number genes assessed after filtering was 5037. (B) Mean absolute error between -log10 p-values of association obtained for high-coverage lead eQTLs and those obtained in each dataset for the same set of variants. All high coverage lead eQTLs (that is a variant for each of the 16894 genes) were considered here, regardless of significance level. The scatterplots detail the -log10 p-values used to calculate the mean absolute errors for several relevant low-coverages and SNP arrays.

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Rubinacci, S., Ribeiro, D.M., Hofmeister, R.J. et al. Efficient phasing and imputation of low-coverage sequencing data using large reference panels. Nat Genet 53, 120–126 (2021).

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