ChipQCreport coverage plot failure?
0
0
Entering edit mode
@tapioenvall-16866
Last seen 6.3 years ago

Hi,

I'm quality-checking the mouse ChIP-seq data with ChIPQC, but for some reason the coverage plot behaves weirdly. After this, I ran the samples by chromosomes and found out it is chr Y - which has much less reads than others - which is causing the trouble. To demonstrate, here are coverage plots with all the chromosomes, with all but chr Y, and with chr Y only.

This is all the data (chr Y included)

https://www.dropbox.com/s/m1e2dzzgqlddumf/with_Y.png?dl=0

This is data without chr Y

https://www.dropbox.com/s/rm4qcou0uuwd030/without_Y.png?dl=0

And this is chr Y itself.

https://www.dropbox.com/s/jsbzjsxovbmbwc5/Y.png?dl=0

Any ideas what is going on, is there something wrong with the data / program?

 

This is my code, (producing the reports in the same order as are the figures):

library("ChIPQC")
samples <- read.csv("samples_ChIPQC", stringsAsFactors = FALSE, header = TRUE)

exampleExp = ChIPQC(samples, chromosomes = NULL)
ChIPQCreport(exampleExp, reportFolder = "bc_all")

#"chromosomes" just lists the chromosomes, Y being the last, 21th
chrs <- (read.table("chromosomes", stringsAsFactors = FALSE))[,1]
exampleExp = ChIPQC(samples, chromosomes = chrs[-21])
ChIPQCreport(exampleExp, reportFolder = "bc_without_Y")
exampleExp = ChIPQC(samples, chromosomes = "chrY")
ChIPQCreport(exampleExp, reportFolder = "bc_chrY")

This is how 'samples' look like.

> samples
  SampleID Tissue    bamReads ControlID  bamControl        Peaks PeakCaller
1       S1     T1 s_1_001.bam   S1_ctrl S1_ctrl.bam S1_peaks.bed       MACS
2       S2     T2 s_2_001.bam   S2_ctrl S2_ctrl.bam S2_peaks.bed       MACS

sessionInfo()

> sessionInfo()
R version 3.5.0 (2018-04-23)
Platform: x86_64-apple-darwin15.6.0 (64-bit)
Running under: macOS High Sierra 10.13.2

Matrix products: default
BLAS: /System/Library/Frameworks/Accelerate.framework/Versions/A/Frameworks/vecLib.framework/Versions/A/libBLAS.dylib
LAPACK: /Library/Frameworks/R.framework/Versions/3.5/Resources/lib/libRlapack.dylib

locale:
[1] en_US.UTF-8/en_US.UTF-8/en_US.UTF-8/C/en_US.UTF-8/en_US.UTF-8

attached base packages:
[1] parallel  stats4    stats     graphics  grDevices utils     datasets  methods   base     

other attached packages:
 [1] ChIPQC_1.16.0               DiffBind_2.8.0              SummarizedExperiment_1.10.1 DelayedArray_0.6.2          BiocParallel_1.14.2         matrixStats_0.54.0         
 [7] Biobase_2.40.0              GenomicRanges_1.32.6        GenomeInfoDb_1.16.0         IRanges_2.14.10             S4Vectors_0.18.3            BiocGenerics_0.26.0        
[13] ggplot2_3.0.0              

loaded via a namespace (and not attached):
  [1] amap_0.8-16                               colorspace_1.3-2                          rjson_0.2.20                             
  [4] hwriter_1.3.2                             XVector_0.20.0                            base64enc_0.1-3                          
  [7] rstudioapi_0.7                            ggrepel_0.8.0                             bit64_0.9-7                              
 [10] AnnotationDbi_1.42.1                      splines_3.5.0                             TxDb.Rnorvegicus.UCSC.rn4.ensGene_3.2.2  
 [13] Nozzle.R1_1.1-1                           Rsamtools_1.32.2                          annotate_1.58.0                          
 [16] GO.db_3.6.0                               pheatmap_1.0.10                           graph_1.58.0                             
 [19] TxDb.Hsapiens.UCSC.hg18.knownGene_3.2.2   compiler_3.5.0                            httr_1.3.1                               
 [22] GOstats_2.46.0                            backports_1.1.2                           assertthat_0.2.0                         
 [25] Matrix_1.2-14                             lazyeval_0.2.1                            TxDb.Hsapiens.UCSC.hg19.knownGene_3.2.2  
 [28] limma_3.36.2                              prettyunits_1.0.2                         tools_3.5.0                              
 [31] bindrcpp_0.2.2                            gtable_0.2.0                              glue_1.3.0                               
 [34] GenomeInfoDbData_1.1.0                    Category_2.46.0                           reshape2_1.4.3                           
 [37] systemPipeR_1.14.0                        dplyr_0.7.6                               ShortRead_1.38.0                         
 [40] Rcpp_0.12.18                              TxDb.Dmelanogaster.UCSC.dm3.ensGene_3.2.2 TxDb.Mmusculus.UCSC.mm9.knownGene_3.2.2  
 [43] Biostrings_2.48.0                         gdata_2.18.0                              rtracklayer_1.40.3                       
 [46] TxDb.Mmusculus.UCSC.mm10.knownGene_3.4.0  stringr_1.3.1                             gtools_3.8.1                             
 [49] XML_3.98-1.13                             edgeR_3.22.3                              zlibbioc_1.26.0                          
 [52] scales_0.5.0                              hms_0.4.2                                 RBGL_1.56.0                              
 [55] RColorBrewer_1.1-2                        BBmisc_1.11                               memoise_1.1.0                            
 [58] biomaRt_2.36.1                            latticeExtra_0.6-28                       stringi_1.2.4                            
 [61] RSQLite_2.1.1                             genefilter_1.62.0                         checkmate_1.8.5                          
 [64] GenomicFeatures_1.32.0                    caTools_1.17.1.1                          chipseq_1.30.0                           
 [67] rlang_0.2.1                               pkgconfig_2.0.1                           BatchJobs_1.7                            
 [70] bitops_1.0-6                              TxDb.Celegans.UCSC.ce6.ensGene_3.2.2      lattice_0.20-35                          
 [73] purrr_0.2.5                               bindr_0.1.1                               labeling_0.3                             
 [76] GenomicAlignments_1.16.0                  bit_1.1-14                                tidyselect_0.2.4                         
 [79] GSEABase_1.42.0                           AnnotationForge_1.22.1                    plyr_1.8.4                               
 [82] magrittr_1.5                              sendmailR_1.2-1                           R6_2.2.2                                 
 [85] gplots_3.0.1                              DBI_1.0.0                                 pillar_1.3.0                             
 [88] withr_2.1.2                               survival_2.42-6                           RCurl_1.95-4.11                          
 [91] tibble_1.4.2                              crayon_1.3.4                              KernSmooth_2.23-15                       
 [94] progress_1.2.0                            locfit_1.5-9.1                            grid_3.5.0                               
 [97] data.table_1.11.4                         blob_1.1.1                                Rgraphviz_2.24.0                         
[100] digest_0.6.15                             xtable_1.8-2                              brew_1.0-6                               
[103] munsell_0.5.0     
ChipQC chipqcreport • 1.1k views
ADD COMMENT
0
Entering edit mode

hi,

Would you be able to share the BAM file with me on Box (tc.infomatics@gmail.com) so I can try and find the source of this?

best,

tom

ADD REPLY
0
Entering edit mode

Hi!

Unfortunately the data is not public and I can not share it.

However, it may have something to do with how chromosomes are combined: I took two non-problematic chromosomes, and started to down-sample the other one, and ran ChIPQC for the combined file. Some "cracks" do appear, though not as drastic as in the figures I linked before. I assume that the coverage plots should approach that of the intact chromosome when inputting less and less reads from the other chromosome.

If I find time I try to reproduce this with some public data.

Best,

Tapio

ADD REPLY

Login before adding your answer.

Traffic: 577 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6