Entering edit mode
I got the following error while trying to create the initiall dba object:
> tamoxifen <- dba.count(tamoxifen)
Computing summits...
Error: Some read files could not be accessed. See warnings for details.
In addition: There were 12 warnings (use warnings() to see them)
> warnings()
Warning messages:
1: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201074_S18_L000/A006200317_201074_S18_L000.bam not accessible
2: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201076_S19_L000/A006200317_201076_S19_L000.bam not accessible
3: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201078_S20_L000/A006200317_201078_S20_L000.bam not accessible
4: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201080_S21_L000/A006200317_201080_S21_L000.bam not accessible
5: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201082_S22_L000/A006200317_201082_S22_L000.bam not accessible
6: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201084_S23_L000/A006200317_201084_S23_L000.bam not accessible
7: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201086_S24_L000/A006200317_201086_S24_L000.bam not accessible
8: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201088_S25_L000/A006200317_201088_S25_L000.bam not accessible
9: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201090_S26_L000/A006200317_201090_S26_L000.bam not accessible
10: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201092_S27_L000/A006200317_201092_S27_L000.bam not accessible
11: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201094_S28_L000/A006200317_201094_S28_L000.bam not accessible
12: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201096_S29_L000/A006200317_201096_S29_L000.bam not accessible
the files are there and using the file.exists I get TRUE at the test. here is a complete code:
read the csv file, I used the sep = \t due to the file being tab delimeted
samples<- read.csv("diffbind.csv",sep = "\t")
> print(samples)
SampleID Factor Condition Replicate
1 201074 H3K36me3 control 1
2 201076 H3K36me3 control 2
3 201078 H3K36me3 control 3
4 201080 H3K36me3 AA5 1
5 201082 H3K36me3 AA5 2
6 201084 H3K36me3 AA5 3
7 201086 H4K16ac control 1
8 201088 H4K16ac control 2
9 201090 H4K16ac control 3
10 201092 H4K16ac AA5 1
11 201094 H4K16ac AA5 2
12 201096 H4K16ac AA5 3
bamReads
1 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201074_S18_L000/A006200317_201074_S18_L000.bam
2 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201076_S19_L000/A006200317_201076_S19_L000.bam
3 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201078_S20_L000/A006200317_201078_S20_L000.bam
4 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201080_S21_L000/A006200317_201080_S21_L000.bam
5 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201082_S22_L000/A006200317_201082_S22_L000.bam
6 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201084_S23_L000/A006200317_201084_S23_L000.bam
7 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201086_S24_L000/A006200317_201086_S24_L000.bam
8 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201088_S25_L000/A006200317_201088_S25_L000.bam
9 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201090_S26_L000/A006200317_201090_S26_L000.bam
10 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201092_S27_L000/A006200317_201092_S27_L000.bam
11 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201094_S28_L000/A006200317_201094_S28_L000.bam
12 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201096_S29_L000/A006200317_201096_S29_L000.bam
Peaks
1 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201074_S18_L000/peakcalling/seacr/A006200317_201074_S18_L000_treat.stringent.sort.bed
2 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201076_S19_L000/peakcalling/seacr/A006200317_201076_S19_L000_treat.stringent.sort.bed
3 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201078_S20_L000/peakcalling/seacr/A006200317_201078_S20_L000_treat.stringent.sort.bed
4 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201080_S21_L000/peakcalling/seacr/A006200317_201080_S21_L000_treat.stringent.sort.bed
5 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201082_S22_L000/peakcalling/seacr/A006200317_201082_S22_L000_treat.stringent.sort.bed
6 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201084_S23_L000/peakcalling/seacr/A006200317_201084_S23_L000_treat.stringent.sort.bed
7 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201086_S24_L000/peakcalling/seacr/A006200317_201086_S24_L000_treat.stringent.sort.bed
8 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201088_S25_L000/peakcalling/seacr/A006200317_201088_S25_L000_treat.stringent.sort.bed
9 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201090_S26_L000/peakcalling/seacr/A006200317_201090_S26_L000_treat.stringent.sort.bed
10 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201092_S27_L000/peakcalling/seacr/A006200317_201092_S27_L000_treat.stringent.sort.bed
11 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201094_S28_L000/peakcalling/seacr/A006200317_201094_S28_L000_treat.stringent.sort.bed
12 Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201096_S29_L000/peakcalling/seacr/A006200317_201096_S29_L000_treat.stringent.sort.bed
PeakCaller scorecol
1 bed 3
2 bed 3
3 bed 3
4 bed 3
5 bed 3
6 bed 3
7 bed 3
8 bed 3
9 bed 3
10 bed 3
11 bed 3
12 bed 3
so the created matrix looks good. lets create the dba object now:
> tamoxifen <- dba(sampleSheet=samples)
201074 H3K36me3 control 1 bed
201076 H3K36me3 control 2 bed
201078 H3K36me3 control 3 bed
201080 H3K36me3 AA5 1 bed
201082 H3K36me3 AA5 2 bed
201084 H3K36me3 AA5 3 bed
201086 H4K16ac control 1 bed
201088 H4K16ac control 2 bed
201090 H4K16ac control 3 bed
201092 H4K16ac AA5 1 bed
201094 H4K16ac AA5 2 bed
201096 H4K16ac AA5 3 bed
> tamoxifen
12 Samples, 3504 sites in matrix (5003 total):
ID Factor Condition Replicate Intervals
1 201074 H3K36me3 control 1 3117
2 201076 H3K36me3 control 2 3042
3 201078 H3K36me3 control 3 2888
4 201080 H3K36me3 AA5 1 3245
5 201082 H3K36me3 AA5 2 3014
6 201084 H3K36me3 AA5 3 3278
7 201086 H4K16ac control 1 114
8 201088 H4K16ac control 2 117
9 201090 H4K16ac control 3 171
10 201092 H4K16ac AA5 1 340
11 201094 H4K16ac AA5 2 282
12 201096 H4K16ac AA5 3 249
this also looks good. I am checking if R in windows 11 can actually find the files:
> file.exists(path = samples$bamReads)
[1] TRUE TRUE TRUE TRUE TRUE TRUE TRUE TRUE TRUE TRUE TRUE TRUE
> file.exists(path = samples$Peaks)
[1] TRUE TRUE TRUE TRUE TRUE TRUE TRUE TRUE TRUE TRUE TRUE TRUE
So all looks ready to do the count
> tamoxifen <- dba.count(tamoxifen)
Computing summits...
Error: Some read files could not be accessed. See warnings for details.
In addition: There were 12 warnings (use warnings() to see them)
> warnings()
Warning messages:
1: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201074_S18_L000/A006200317_201074_S18_L000.bam not accessible
2: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201076_S19_L000/A006200317_201076_S19_L000.bam not accessible
3: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201078_S20_L000/A006200317_201078_S20_L000.bam not accessible
4: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201080_S21_L000/A006200317_201080_S21_L000.bam not accessible
5: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201082_S22_L000/A006200317_201082_S22_L000.bam not accessible
6: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201084_S23_L000/A006200317_201084_S23_L000.bam not accessible
7: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201086_S24_L000/A006200317_201086_S24_L000.bam not accessible
8: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201088_S25_L000/A006200317_201088_S25_L000.bam not accessible
9: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201090_S26_L000/A006200317_201090_S26_L000.bam not accessible
10: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201092_S27_L000/A006200317_201092_S27_L000.bam not accessible
11: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201094_S28_L000/A006200317_201094_S28_L000.bam not accessible
12: Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201096_S29_L000/A006200317_201096_S29_L000.bam not accessible
I have no idea how to solve this any clues? SESSION INFO
sessionInfo( )
R version 4.2.1 (2022-06-23 ucrt)
Platform: x86_64-w64-mingw32/x64 (64-bit)
Running under: Windows 10 x64 (build 22621)
Matrix products: default
locale:
[1] LC_COLLATE=English_United Kingdom.utf8 LC_CTYPE=English_United Kingdom.utf8 LC_MONETARY=English_United Kingdom.utf8
[4] LC_NUMERIC=C LC_TIME=English_United Kingdom.utf8
attached base packages:
[1] stats4 stats graphics grDevices utils datasets methods base
other attached packages:
[1] DiffBind_3.8.4 SummarizedExperiment_1.28.0 Biobase_2.58.0 MatrixGenerics_1.10.0
[5] matrixStats_0.63.0 GenomicRanges_1.50.2 GenomeInfoDb_1.34.9 IRanges_2.32.0
[9] S4Vectors_0.36.2 BiocGenerics_0.44.0
loaded via a namespace (and not attached):
[1] bitops_1.0-7 RColorBrewer_1.1-3 numDeriv_2016.8-1.1 tools_4.2.1 utf8_1.2.3
[6] R6_2.5.1 irlba_2.3.5.1 KernSmooth_2.23-20 DBI_1.1.3 colorspace_2.1-0
[11] apeglm_1.20.0 tidyselect_1.2.0 compiler_4.2.1 cli_3.6.0 DelayedArray_0.24.0
[16] rtracklayer_1.58.0 caTools_1.18.2 scales_1.2.1 SQUAREM_2021.1 mvtnorm_1.1-3
[21] mixsqp_0.3-48 stringr_1.5.0 digest_0.6.31 Rsamtools_2.14.0 XVector_0.38.0
[26] jpeg_0.1-10 pkgconfig_2.0.3 htmltools_0.5.4 fastmap_1.1.1 invgamma_1.1
[31] bbmle_1.0.25 limma_3.54.2 BSgenome_1.66.3 htmlwidgets_1.6.2 rlang_1.1.0
[36] rstudioapi_0.14 BiocIO_1.8.0 generics_0.1.3 hwriter_1.3.2.1 BiocParallel_1.32.6
[41] gtools_3.9.4 dplyr_1.1.2 RCurl_1.98-1.12 magrittr_2.0.3 GenomeInfoDbData_1.2.9
[46] interp_1.1-4 Matrix_1.5-3 Rcpp_1.0.10 munsell_0.5.0 fansi_1.0.4
[51] lifecycle_1.0.3 stringi_1.7.12 yaml_2.3.7 MASS_7.3-58.3 zlibbioc_1.44.0
[56] gplots_3.1.3 plyr_1.8.8 grid_4.2.1 parallel_4.2.1 ggrepel_0.9.3
[61] bdsmatrix_1.3-6 crayon_1.5.2 deldir_1.0-9 lattice_0.21-8 Biostrings_2.66.0
[66] locfit_1.5-9.7 pillar_1.9.0 rjson_0.2.21 systemPipeR_2.4.0 codetools_0.2-18
[71] XML_3.99-0.14 glue_1.6.2 ShortRead_1.56.1 GreyListChIP_1.30.0 latticeExtra_0.6-30
[76] BiocManager_1.30.20 png_0.1-8 vctrs_0.6.0 gtable_0.3.3 amap_0.8-19
[81] ashr_2.2-54 ggplot2_3.4.2 emdbook_1.3.12 restfulr_0.0.15 coda_0.19-4
[86] truncnorm_1.0-9 tibble_3.2.1 GenomicAlignments_1.34.1
>
you hit the spot. But this is a weird issue. Using windows file manager I got, my username as owner. Using WSL2 I am seeing the owner is root and group is again root. permission though are read and write for all. interesting is that I got in the same directory the peaks file that can be accessed without issues, but
file.access()
gives me a-1
. ```R file.access(samples$bamReads, 4) Z:/Shared folder/Shared data/DNA/NGS_AP01_cfrezza_A006200317/A006200317_201074_S18_L000/A006200317_201074_S18_L000.bam -1The peak file has been nicely read:
That is weird. No idea why the bed file would be accessible but not the bam. I assume Z: is a samba share? Over the years I have seen people having problems reading files off a samba share on Windows, so maybe it's just something having to do with that. As an example, sometimes people try to put their R library dir on a shared drive and that usually ends in tears. The normal prescription is to tell people not to do that, but moving a bunch of bam files onto your Windows box might not be something you can do.
Sorry I can't be more helpful.
No worries, moved those locally and that error disaeared. Must to be something woth samba servers. Any way. Now I got another problem and I will open a new question. Thank you James!