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lauren.fitch
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@laurenfitch-11575
Last seen 4.7 years ago
I have run DESeq2 successfully on my dataset with several different contrasts; however, when I try to run this one I get the following error:
> dds <- DESeqDataSetFromMatrix( + countData = countdata.strict, + colData = samples.strict, + design = ~ group + ) converting counts to integer mode > dds <- DESeq(dds) estimating size factors estimating dispersions gene-wise dispersion estimates mean-dispersion relationship final dispersion estimates fitting model and testing -- replacing outliers and refitting for 1279 genes -- DESeq argument 'minReplicatesForReplace' = 7 -- original counts are preserved in counts(dds) estimating dispersions fitting model and testing > res <- results(dds, contrast = c("group", "HDGSE85", "HDGSE86")) Error in cleanContrast(object, contrast, expanded = isExpanded, listValues = listValues, : groupHDGSE85 and groupHDGSE86 are expected to be in resultsNames(object) > resultsNames(dds) [1] "Intercept" "groupHDGSE86" "groupHDGSE87" "groupHDSGSE85" >
"group" in this case indicates sequencing batch. I have run a contrast using "state" already with no problem. Here's what my sample data looks like:
> samples.strict
group genealine state sex filter.strict filter.relaxed replicate observationid EffectiveAlignmentCount
G002.R1.CTL HDSGSE85 G002 CTL M FALSE FALSE 1 G002_CD140_a 151040648
G002.R2.CTL HDSGSE85 G002 CTL M FALSE FALSE 2 G002_CD140_b 142144646
G002.R3.CTL HDSGSE85 G002 CTL M FALSE FALSE 3 G002_CD140_c 128589118
G002.R4.CTL HDSGSE85 G002 CTL M FALSE FALSE 4 G002_CD140_d 152415461
G002.R5.CTL HDSGSE85 G002 CTL M FALSE FALSE 5 G002_CD140_e 135059579
G002.R6.CTL HDSGSE85 G002 CTL M FALSE FALSE 6 G002_CD140_f 121105092
G017.R1.HD HDGSE86 G017 HD M FALSE FALSE 1 SG17a 44082945
G017.R2.HD HDGSE86 G017 HD M FALSE FALSE 2 SG17b 43321404
G017.R3.HD HDGSE86 G017 HD M FALSE FALSE 3 SG17c 39897764
G017.R4.HD HDGSE86 G017 HD M FALSE FALSE 4 SG17e 55759593
G017.R5.HD HDGSE86 G017 HD M FALSE FALSE 5 SG17f 40662631
G018.R1.HD HDGSE86 G018 HD F FALSE FALSE 1 SG18a 47275799
G018.R3.HD HDGSE86 G018 HD F FALSE FALSE 3 SG18c 44562212
G018.R4.HD HDGSE86 G018 HD F FALSE FALSE 4 SG18d 48443633
G018.R5.HD HDGSE86 G018 HD F FALSE FALSE 5 SG18f 47221901
G019.R6.CTL HDGSE87 G019 CTL F FALSE FALSE 6 SG-G19d 23390368
G020.R3.HD HDGSE87 G020 HD F FALSE FALSE 3 SG-G20a 20107140
>
> sessionInfo()
R version 3.3.1 (2016-06-21) Platform: x86_64-w64-mingw32/x64 (64-bit) Running under: Windows 7 x64 (build 7601) Service Pack 1 locale: [1] LC_COLLATE=English_United States.1252 LC_CTYPE=English_United States.1252 LC_MONETARY=English_United States.1252 [4] LC_NUMERIC=C LC_TIME=English_United States.1252 attached base packages: [1] parallel stats4 stats graphics grDevices utils datasets methods base other attached packages: [1] calibrate_1.7.2 MASS_7.3-45 genefilter_1.56.0 RColorBrewer_1.1-2 [5] gplots_3.0.1 DESeq2_1.14.0 SummarizedExperiment_1.4.0 Biobase_2.34.0 [9] GenomicRanges_1.26.1 GenomeInfoDb_1.10.0 IRanges_2.8.0 S4Vectors_0.12.0 [13] BiocGenerics_0.20.0 loaded via a namespace (and not attached): [1] Rcpp_0.12.7 plyr_1.8.4 XVector_0.14.0 bitops_1.0-6 tools_3.3.1 [6] zlibbioc_1.20.0 rpart_4.1-10 RSQLite_1.0.0 annotate_1.52.0 gtable_0.2.0 [11] lattice_0.20-33 Matrix_1.2-6 DBI_0.5-1 gridExtra_2.2.1 cluster_2.0.4 [16] caTools_1.17.1 gtools_3.5.0 locfit_1.5-9.1 grid_3.3.1 nnet_7.3-12 [21] data.table_1.9.6 AnnotationDbi_1.36.0 XML_3.98-1.4 survival_2.39-5 BiocParallel_1.8.0 [26] foreign_0.8-67 gdata_2.17.0 latticeExtra_0.6-28 Formula_1.2-1 geneplotter_1.52.0 [31] ggplot2_2.1.0 Hmisc_3.17-4 scales_0.4.0 splines_3.3.1 colorspace_1.2-7 [36] xtable_1.8-2 KernSmooth_2.23-15 acepack_1.4.0 RCurl_1.95-4.8 munsell_0.4.3 [41] chron_2.3-47
I was able to run the contrast successfully using this code. I'm still not sure why the original doesn't work, though.