Hi,

I'm analyzing label-free proteomics data with DEP, but have run into an error of having duplicate row names.

Here's what I have done: I have generated unique identifiers as indicated in the DEP tutorial:

> head(data)
# A tibble: 6 x 26
  Protein.IDs    Gene.names LNP34_i2_1 LNP34_i2_2 LNP34_i2_3 LNP34_n_1 LNP34_n_2
  <chr>          <chr>           <dbl>      <dbl>      <dbl>     <dbl>     <dbl>
1 P08226;A0A1B0~ Apoe          2.93e10    6.03e 9    2.79e10   3.60e10   2.63e10
2 P07724         Alb           6.36e 9    1.20e10    5.84e 9   6.28e 9   3.17e 9
3 A8DUK4;E9Q223~ Hbb-bs        2.20e 9    2.70e 9    2.85e 9   5.09e 9   2.13e 9
4 Q91VB8;P01942~ Hba-a1        1.51e 9    2.84e 9    2.00e 9   2.86e 9   2.12e 9
5 A0A075B5P6;A0~ Ighm          1.82e 9    1.28e 9    1.77e 9   4.15e 8   2.44e 9
6 Q921I1;F7BAE9~ Tf            5.06e 8    1.59e 9    5.44e 8   4.20e 8   2.53e 8
# ... with 19 more variables: LNP34_n_3 <dbl>, LNP35_i2_1 <dbl>,
#   LNP35_i2_2 <dbl>, LNP35_i2_3 <dbl>, LNP35_n_1 <dbl>, LNP35_n_2 <dbl>,
#   LNP35_n_3 <dbl>, LNP36_i2_1 <dbl>, LNP36_i2_2 <dbl>, LNP36_i2_3 <dbl>,
#   LNP36_n_1 <dbl>, LNP36_n_2 <dbl>, LNP36_n_3 <dbl>, LNP37_i2_1 <dbl>,
#   LNP37_i2_2 <dbl>, LNP37_i2_3 <dbl>, LNP37_n_1 <dbl>, LNP37_n_2 <dbl>,
#   LNP37_n_3 <dbl>

> data$Gene.names %>% duplicated() %>% any()
[1] TRUE

> data %>% group_by(Gene.names) %>% summarize(frequency = n()) %>% arrange(desc(frequency)) %>% filter(frequency > 1)
# A tibble: 6 x 2
  Gene.names frequency
  <chr>          <int>
1 _                 19
2 H2-K1              2
3 Itih4              2
4 Kng1               2
5 Sptb               2
6 Tpm3               2

> data_unique <- make_unique(data, "Gene.names", "Protein.IDs", delim = ";")

> data_unique$name %>% duplicated() %>% any()
[1] FALSE

....and would now like to generate a SummarizedExperiment by using my own experimental design. Yet, receive an error on duplicate row names and non-unique values

> experimental_design
        label condition LNPcondition replicate
1  LNP34_i2_1     LNP34     LNP34_i2         1
2  LNP34_i2_2     LNP34     LNP34_i2         2
3  LNP34_i2_3     LNP34     LNP34_i2         3
4   LNP34_n_1     LNP34      LNP34_n         1
5   LNP34_n_2     LNP34      LNP34_n         2
6   LNP34_n_3     LNP34      LNP34_n         3
7  LNP35_i2_1     LNP35     LNP35_i2         1
8  LNP35_i2_2     LNP35     LNP35_i2         2
9  LNP35_i2_3     LNP35     LNP35_i2         3
10  LNP35_n_1     LNP35      LNP35_n         1
11  LNP35_n_2     LNP35      LNP35_n         2
12  LNP35_n_3     LNP35      LNP35_n         3
13 LNP36_i2_1     LNP36     LNP36_i2         1
14 LNP36_i2_2     LNP36     LNP36_i2         2
15 LNP36_i2_3     LNP36     LNP36_i2         3
16  LNP36_n_1     LNP36      LNP36_n         1
17  LNP36_n_2     LNP36      LNP36_n         2
18  LNP36_n_3     LNP36      LNP36_n         3
19 LNP37_i2_1     LNP37     LNP37_i2         1
20 LNP37_i2_2     LNP37     LNP37_i2         2
21 LNP37_i2_3     LNP37     LNP37_i2         3
22  LNP37_n_1     LNP37      LNP37_n         1
23  LNP37_n_2     LNP37      LNP37_n         2
24  LNP37_n_3     LNP37      LNP37_n         3

> LFQ_columns <- grep("^LNP", colnames(data_unique))
> data_se <- make_se(data_unique, LFQ_columns, experimental_design)
Error in `.rowNamesDF<-`(x, value = value) : 
  duplicate 'row.names' are not allowed
In addition: Warning message:
non-unique values when setting 'row.names': ‘LNP34_1’, ‘LNP34_2’, ‘LNP34_3’, ‘LNP35_1’, ‘LNP35_2’, ‘LNP35_3’, ‘LNP36_1’, ‘LNP36_2’, ‘LNP36_3’, ‘LNP37_1’, ‘LNP37_2’, ‘LNP37_3’

I'm not sure where the error comes from (would guess that from the "condition" column of experimental_design). Though, when checking, no duplicates are found:

> any(duplicated(rownames(experimental_design)))
[1] FALSE
> any(duplicated(rownames(data_unique)))
[1] FALSE
> any(duplicated(rownames(LFQ_columns)))
[1] FALSE
> any(duplicated(colnames(experimental_design)))
[1] FALSE
> any(duplicated(colnames(data_unique)))
[1] FALSE
> any(duplicated(colnames(LFQ_columns)))

Can someone please help me on this? I have a limited knowledge in R, hence details are highly appreciated.

Sessioninfo below. Thanks!

> sessionInfo()
R version 4.0.2 (2020-06-22)
Platform: x86_64-w64-mingw32/x64 (64-bit)
Running under: Windows 10 x64 (build 18363)

Matrix products: default

locale:
[1] LC_COLLATE=English_United Kingdom.1252 
[2] LC_CTYPE=English_United Kingdom.1252   
[3] LC_MONETARY=English_United Kingdom.1252
[4] LC_NUMERIC=C                           
[5] LC_TIME=English_United Kingdom.1252    

attached base packages:
[1] stats     graphics  grDevices utils     datasets  methods   base     

other attached packages:
[1] dplyr_1.0.5  DEP_1.10.0   readxl_1.3.1 readr_1.4.0 

loaded via a namespace (and not attached):
  [1] ProtGenerics_1.20.0         bitops_1.0-6               
  [3] matrixStats_0.58.0          doParallel_1.0.16          
  [5] RColorBrewer_1.1-2          GenomeInfoDb_1.24.2        
  [7] MSnbase_2.14.2              tools_4.0.2                
  [9] DT_0.17                     utf8_1.2.1                 
 [11] R6_2.5.0                    affyio_1.58.0              
 [13] tmvtnorm_1.4-10             BiocGenerics_0.34.0        
 [15] colorspace_2.0-0            GetoptLong_1.0.5           
 [17] tidyselect_1.1.0            compiler_4.0.2             
 [19] preprocessCore_1.50.0       cli_2.4.0                  
 [21] Biobase_2.48.0              DelayedArray_0.14.1        
 [23] sandwich_3.0-0              scales_1.1.1               
 [25] mvtnorm_1.1-1               affy_1.66.0                
 [27] digest_0.6.27               XVector_0.28.0             
 [29] htmltools_0.5.1.1           pkgconfig_2.0.3            
 [31] fastmap_1.1.0               limma_3.44.3               
 [33] htmlwidgets_1.5.3           rlang_0.4.10               
 [35] GlobalOptions_0.1.2         rstudioapi_0.13            
 [37] impute_1.62.0               shiny_1.6.0                
 [39] shape_1.4.5                 generics_0.1.0             
 [41] zoo_1.8-9                   mzID_1.26.0                
 [43] BiocParallel_1.22.0         RCurl_1.98-1.3             
 [45] magrittr_2.0.1              GenomeInfoDbData_1.2.3     
 [47] MALDIquant_1.19.3           Matrix_1.2-18              
 [49] Rcpp_1.0.6                  munsell_0.5.0              
 [51] S4Vectors_0.26.1            fansi_0.4.2                
 [53] imputeLCMD_2.0              lifecycle_1.0.0            
 [55] vsn_3.56.0                  MASS_7.3-51.6              
 [57] SummarizedExperiment_1.18.2 zlibbioc_1.34.0            
 [59] plyr_1.8.6                  grid_4.0.2                 
 [61] promises_1.2.0.1            parallel_4.0.2             
 [63] shinydashboard_0.7.1        crayon_1.4.1               
 [65] lattice_0.20-41             circlize_0.4.12            
 [67] hms_1.0.0                   mzR_2.22.0                 
 [69] ComplexHeatmap_2.4.3        pillar_1.5.1               
 [71] GenomicRanges_1.40.0        rjson_0.2.20               
 [73] codetools_0.2-16            stats4_4.0.2               
 [75] XML_3.99-0.6                glue_1.4.2                 
 [77] pcaMethods_1.80.0           BiocManager_1.30.12        
 [79] httpuv_1.5.5                png_0.1-7                  
 [81] vctrs_0.3.7                 foreach_1.5.1              
 [83] cellranger_1.1.0            tidyr_1.1.3                
 [85] gtable_0.3.0                purrr_0.3.4                
 [87] norm_1.0-9.5                clue_0.3-58                
 [89] assertthat_0.2.1            ggplot2_3.3.3              
 [91] xfun_0.22                   mime_0.10                  
 [93] xtable_1.8-4                later_1.1.0.1              
 [95] ncdf4_1.17                  tibble_3.1.0               
 [97] iterators_1.0.13            gmm_1.6-6                  
 [99] tinytex_0.31                IRanges_2.22.2             
[101] cluster_2.1.0               ellipsis_0.3.1