Hi,
I have a 2 factor experiment (2 mutations:A,B), with 2 levels (+/-) in each factor. All together there 4 conditions: wt, A, B, and A+B , with two replicates for each condition. I tried to use LRT test to isolate genes that are correspond to the mutations without interaction. the full model I used is ~A+B+A:B and the reduced is ~A:B I received the following error:
Error in nbinomLRT(object, full = full, reduced = reduced, quiet = quiet, :
less than one degree of freedom, perhaps full and reduced models are not in the correct order
why are the degrees of freedom less than 1? Below I pasted the sessionInfo output. Thanks in advance.
Guy
R version 4.0.2 (2020-06-22)
Platform: x86_64-w64-mingw32/x64 (64-bit)
Running under: Windows 10 x64 (build 18362)
Matrix products: default
Random number generation:
RNG: Mersenne-Twister
Normal: Inversion
Sample: Rounding
locale:
[1] LC_COLLATE=English_United States.1252
[2] LC_CTYPE=English_United States.1252
[3] LC_MONETARY=English_United States.1252
[4] LC_NUMERIC=C
[5] LC_TIME=English_United States.1252
attached base packages:
[1] grid parallel stats4 stats graphics grDevices
[7] utils datasets methods base
other attached packages:
[1] ggplotify_0.0.5 gridExtra_2.3
[3] dplyr_1.0.0 DESeq2_1.28.1
[5] SummarizedExperiment_1.18.1 DelayedArray_0.14.0
[7] matrixStats_0.56.0 Biobase_2.48.0
[9] GenomicRanges_1.40.0 GenomeInfoDb_1.24.2
[11] IRanges_2.22.2 S4Vectors_0.26.1
[13] BiocGenerics_0.34.0 ggplot2_3.3.2
[15] readxl_1.3.1 data.table_1.12.8
loaded via a namespace (and not attached):
[1] Rcpp_1.0.4.6 locfit_1.5-9.4
[3] lattice_0.20-41 assertthat_0.2.1
[5] digest_0.6.25 R6_2.4.1
[7] cellranger_1.1.0 RSQLite_2.2.0
[9] pillar_1.4.4 zlibbioc_1.34.0
[11] rlang_0.4.6 rstudioapi_0.11
[13] annotate_1.66.0 blob_1.2.1
[15] Matrix_1.2-18 labeling_0.3
[17] splines_4.0.2 BiocParallel_1.22.0
[19] geneplotter_1.66.0 pheatmap_1.0.12
[21] RCurl_1.98-1.2 bit_1.1-15.2
[23] munsell_0.5.0 compiler_4.0.2
[25] pkgconfig_2.0.3 gridGraphics_0.5-0
[27] tidyselect_1.1.0 tibble_3.0.1
[29] GenomeInfoDbData_1.2.3 XML_3.99-0.4
[31] fansi_0.4.1 crayon_1.3.4
[33] withr_2.2.0 bitops_1.0-6
[35] xtable_1.8-4 gtable_0.3.0
[37] lifecycle_0.2.0 DBI_1.1.0
[39] magrittr_1.5 scales_1.1.1
[41] cli_2.0.2 farver_2.0.3
[43] XVector_0.28.0 genefilter_1.70.0
[45] ellipsis_0.3.1 rvcheck_0.1.8
[47] generics_0.0.2 vctrs_0.3.1
[49] RColorBrewer_1.1-2 tools_4.0.2
[51] bit64_0.9-7 glue_1.4.1
[53] purrr_0.3.4 survival_3.1-12
[55] AnnotationDbi_1.50.1 colorspace_1.4-1
[57] BiocManager_1.30.10 memoise_1.1.0
Thanks for the help.
So to find genes affected only by mutation A (and not by mutation B or interaction) should I: 1)test full=~A+B+A:B, reduced=~1 2)test full=~A+B+A:B, reduced=~A+B 3) test full=~A+B, reduced=~B
Select genes significant in 1 and not significant in 2 and 3?
I'd recommend collaborating with a statistician to work out the right statistical design for your experiment. We have a lot of material in the vignette, but beyond that I unfortunately don't have time to provide statistical consultation on the support site.