Last updated: 2019-01-03
workflowr checks: (Click a bullet for more information)Great! Since the R Markdown file has been committed to the Git repository, you know the exact version of the code that produced these results.
Great job! The global environment was empty. Objects defined in the global environment can affect the analysis in your R Markdown file in unknown ways. For reproduciblity it’s best to always run the code in an empty environment.
set.seed(12345)
The command set.seed(12345)
was run prior to running the code in the R Markdown file. Setting a seed ensures that any results that rely on randomness, e.g. subsampling or permutations, are reproducible.
Great job! Recording the operating system, R version, and package versions is critical for reproducibility.
wflow_publish
or wflow_git_commit
). workflowr only checks the R Markdown file, but you know if there are other scripts or data files that it depends on. Below is the status of the Git repository when the results were generated:
Ignored files:
Ignored: .DS_Store
Ignored: .Rhistory
Ignored: .Rproj.user/
Ignored: data/.DS_Store
Ignored: output/.DS_Store
Untracked files:
Untracked: KalistoAbundance18486.txt
Untracked: analysis/DirectionapaQTL.Rmd
Untracked: analysis/ncbiRefSeq_sm.sort.mRNA.bed
Untracked: analysis/snake.config.notes.Rmd
Untracked: analysis/verifyBAM.Rmd
Untracked: code/PeaksToCoverPerReads.py
Untracked: code/strober_pc_pve_heatmap_func.R
Untracked: data/18486.genecov.txt
Untracked: data/APApeaksYL.total.inbrain.bed
Untracked: data/ChromHmmOverlap/
Untracked: data/GM12878.chromHMM.bed
Untracked: data/GM12878.chromHMM.txt
Untracked: data/LianoglouLCL/
Untracked: data/LocusZoom/
Untracked: data/NuclearApaQTLs.txt
Untracked: data/PeakCounts/
Untracked: data/PeaksUsed/
Untracked: data/RNAkalisto/
Untracked: data/TotalApaQTLs.txt
Untracked: data/Totalpeaks_filtered_clean.bed
Untracked: data/UnderstandPeaksQC/
Untracked: data/YL-SP-18486-T-combined-genecov.txt
Untracked: data/YL-SP-18486-T_S9_R1_001-genecov.txt
Untracked: data/apaExamp/
Untracked: data/bedgraph_peaks/
Untracked: data/bin200.5.T.nuccov.bed
Untracked: data/bin200.Anuccov.bed
Untracked: data/bin200.nuccov.bed
Untracked: data/clean_peaks/
Untracked: data/comb_map_stats.csv
Untracked: data/comb_map_stats.xlsx
Untracked: data/comb_map_stats_39ind.csv
Untracked: data/combined_reads_mapped_three_prime_seq.csv
Untracked: data/diff_iso_trans/
Untracked: data/ensemble_to_genename.txt
Untracked: data/example_gene_peakQuant/
Untracked: data/explainProtVar/
Untracked: data/filtered_APApeaks_merged_allchrom_refseqTrans.closest2End.bed
Untracked: data/filtered_APApeaks_merged_allchrom_refseqTrans.closest2End.noties.bed
Untracked: data/first50lines_closest.txt
Untracked: data/gencov.test.csv
Untracked: data/gencov.test.txt
Untracked: data/gencov_zero.test.csv
Untracked: data/gencov_zero.test.txt
Untracked: data/gene_cov/
Untracked: data/joined
Untracked: data/leafcutter/
Untracked: data/merged_combined_YL-SP-threeprimeseq.bg
Untracked: data/mol_overlap/
Untracked: data/mol_pheno/
Untracked: data/nom_QTL/
Untracked: data/nom_QTL_opp/
Untracked: data/nom_QTL_trans/
Untracked: data/nuc6up/
Untracked: data/other_qtls/
Untracked: data/pQTL_otherphen/
Untracked: data/peakPerRefSeqGene/
Untracked: data/perm_QTL/
Untracked: data/perm_QTL_opp/
Untracked: data/perm_QTL_trans/
Untracked: data/perm_QTL_trans_filt/
Untracked: data/reads_mapped_three_prime_seq.csv
Untracked: data/smash.cov.results.bed
Untracked: data/smash.cov.results.csv
Untracked: data/smash.cov.results.txt
Untracked: data/smash_testregion/
Untracked: data/ssFC200.cov.bed
Untracked: data/temp.file1
Untracked: data/temp.file2
Untracked: data/temp.gencov.test.txt
Untracked: data/temp.gencov_zero.test.txt
Untracked: data/threePrimeSeqMetaData.csv
Untracked: output/picard/
Untracked: output/plots/
Untracked: output/qual.fig2.pdf
Unstaged changes:
Modified: analysis/28ind.peak.explore.Rmd
Modified: analysis/InvestigatePeak2GeneAssignment.Rmd
Modified: analysis/apaQTLoverlapGWAS.Rmd
Modified: analysis/cleanupdtseq.internalpriming.Rmd
Modified: analysis/coloc_apaQTLs_protQTLs.Rmd
Modified: analysis/dif.iso.usage.leafcutter.Rmd
Modified: analysis/diff_iso_pipeline.Rmd
Modified: analysis/explainpQTLs.Rmd
Modified: analysis/explore.filters.Rmd
Modified: analysis/flash2mash.Rmd
Modified: analysis/overlapMolQTL.Rmd
Modified: analysis/overlap_qtls.Rmd
Modified: analysis/peakOverlap_oppstrand.Rmd
Modified: analysis/pheno.leaf.comb.Rmd
Modified: analysis/swarmPlots_QTLs.Rmd
Modified: analysis/test.max2.Rmd
Modified: analysis/understandPeaks.Rmd
Modified: code/Snakefile
Note that any generated files, e.g. HTML, png, CSS, etc., are not included in this status report because it is ok for generated content to have uncommitted changes. File | Version | Author | Date | Message |
---|---|---|---|---|
Rmd | c3b3bbb | Briana Mittleman | 2019-01-03 | start covariate correlation with pc analysis |
In this analysis I will look at which collected covariates help explain the variation in the peak data. I am using code from Ben Strobers github, available at https://github.com/BennyStrobes/ipsc_preprocess_pipeline. Specifcially I am looking at the covariate_pc_pve_heatmap function.
library(tidyverse)
── Attaching packages ────────────────────────────────────────────────────────── tidyverse 1.2.1 ──
✔ ggplot2 3.0.0 ✔ purrr 0.2.5
✔ tibble 1.4.2 ✔ dplyr 0.7.6
✔ tidyr 0.8.1 ✔ stringr 1.3.1
✔ readr 1.1.1 ✔ forcats 0.3.0
── Conflicts ───────────────────────────────────────────────────────────── tidyverse_conflicts() ──
✖ dplyr::filter() masks stats::filter()
✖ dplyr::lag() masks stats::lag()
library(workflowr)
This is workflowr version 1.1.1
Run ?workflowr for help getting started
library(cowplot)
Attaching package: 'cowplot'
The following object is masked from 'package:ggplot2':
ggsave
library(reshape2)
Attaching package: 'reshape2'
The following object is masked from 'package:tidyr':
smiths
Load in coverage files:
total_Cov=read.table("../data/PeakCounts/filtered_APApeaks_merged_allchrom_refseqGenes.Transcript_sm_quant.Total_fixed.fc", header=T, stringsAsFactors = F)[,7:45]
nuclear_Cov=read.table("../data/PeakCounts/filtered_APApeaks_merged_allchrom_refseqGenes.Transcript_sm_quant.Nuclear_fixed.fc", header=T, stringsAsFactors = F)[,7:45]
Perform PCA:
Total
pca_tot_peak=prcomp(total_Cov, center=T,scale=T)
pca_tot_df=as.data.frame(pca_tot_peak$rotation) %>% rownames_to_column(var="lib") %>% select(1:11) %>% mutate(line=substr(lib,2,6))
pca_tot_df_fix=bind_cols(line=pca_tot_df[,dim(pca_tot_df)[[2]]],pca_tot_df[,3:dim(pca_tot_df)[[2]]-1])
Nuclear
pca_nuc_peak=prcomp(nuclear_Cov, center=T,scale=T)
pca_nuc_df=as.data.frame(pca_nuc_peak$rotation) %>% rownames_to_column(var="lib") %>% select(1:11) %>% mutate(line=substr(lib,2,6))
pca_nuc_df_fix=bind_cols(line=pca_nuc_df[,dim(pca_nuc_df)[[2]]],pca_nuc_df[,3:dim(pca_nuc_df)[[2]]-1])
Get the line order as a vector
line_order=pca_nuc_df_fix[["line"]]
Load covariate File- filter out lines not yet sequenced and reorder.
covar=read.csv("../data/threePrimeSeqMetaData.csv")[1:78,]
Subset by fraction:
tot_covar=covar %>% filter(fraction=="total") %>% slice(match(line_order, line))
nuc_covar=covar %>% filter(fraction=="nuclear")%>% slice(match(line_order, line))
Subset only a few covariates to try first:
tot_covar_filt=tot_covar %>% select(batch,comb_mapped,Sex, alive_avg, undiluted_avg, cycles)
nuc_covar_filt=nuc_covar %>% select(batch,Sex, alive_avg, undiluted_avg, cycles)
Update Ben’s Function for my data:
covariate_pc_pve_heatmap <- function(pc_df, covariate_df, output_file, title) {
# Load in data
pcs <- pc_df
covs <- covariate_df
# Remove unimportant columns
pcs <- as.matrix(pcs[,2:dim(pcs)[[2]]])
covs <- data.frame(as.matrix(covs[,1:dim(covs)[[2]]]))
# Initialize PVE heatmap
pve_map <- matrix(0, dim(covs)[2], dim(pcs)[2])
colnames(pve_map) <- colnames(pcs)
rownames(pve_map) <- colnames(covs)
# Loop through each PC, COV Pair and take correlation
num_pcs <- dim(pcs)[2]
num_covs <- dim(covs)[2]
for (num_pc in 1:num_pcs) {
for (num_cov in 1:num_covs) {
pc_vec <- pcs[,num_pc]
cov_vec <- covs[,num_cov]
lin_model <- lm(pc_vec ~ cov_vec)
pve_map[num_cov, num_pc] <- summary(lin_model)$adj.r.squared
}
}
pve_map
ord <- hclust( dist(scale(pve_map), method = "euclidean"), method = "ward.D" )$order
melted_mat <- melt(pve_map)
colnames(melted_mat) <- c("Covariate", "PC","PVE")
# Use factors to represent covariate and pc name
melted_mat$Covariate <- factor(melted_mat$Covariate, levels = rownames(pve_map)[ord])
melted_mat$PC <- factor(melted_mat$PC)
if (dim(pcs)[2] == 10) {
levels(melted_mat$PC) <- c(levels(melted_mat$PC)[1],levels(melted_mat$PC)[3:10],levels(melted_mat$PC)[2])
}
if (dim(pcs)[2] == 21) {
levels(melted_mat$PC) <- c(levels(melted_mat$PC)[1],levels(melted_mat$PC)[12],levels(melted_mat$PC)[15:21],levels(melted_mat$PC)[2:11], levels(melted_mat$PC)[13:14])
}
# PLOT!
heatmap <- ggplot(data=melted_mat, aes(x=Covariate, y=PC)) + geom_tile(aes(fill=PVE)) + scale_fill_gradient2(midpoint=-.05, guide="colorbar")
heatmap <- heatmap + theme(text = element_text(size=14), panel.background = element_blank(), axis.text.x = element_text(angle = 90, vjust=.5))
heatmap <- heatmap + labs(y="latent factor", title=title)
# Save File
ggsave(heatmap, file=output_file,width = 19,height=13.5,units="cm")
}
Try it:
Total
covariate_pc_pve_heatmap(pca_tot_df_fix, tot_covar_filt, "../output/plots/TotalCovariatesagainstPCs.39ind.png", "Total Covariates")
Try it: Nuclear
covariate_pc_pve_heatmap(pca_nuc_df_fix, nuc_covar_filt, "../output/plots/NuclearCovariatesagainstPCs.39ind.png", "Nuclear Covariates")
sessionInfo()
R version 3.5.1 (2018-07-02)
Platform: x86_64-apple-darwin15.6.0 (64-bit)
Running under: macOS 10.14.1
Matrix products: default
BLAS: /Library/Frameworks/R.framework/Versions/3.5/Resources/lib/libRblas.0.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] stats graphics grDevices utils datasets methods base
other attached packages:
[1] bindrcpp_0.2.2 reshape2_1.4.3 cowplot_0.9.3 workflowr_1.1.1
[5] forcats_0.3.0 stringr_1.3.1 dplyr_0.7.6 purrr_0.2.5
[9] readr_1.1.1 tidyr_0.8.1 tibble_1.4.2 ggplot2_3.0.0
[13] tidyverse_1.2.1
loaded via a namespace (and not attached):
[1] tidyselect_0.2.4 haven_1.1.2 lattice_0.20-35
[4] colorspace_1.3-2 htmltools_0.3.6 yaml_2.2.0
[7] rlang_0.2.2 R.oo_1.22.0 pillar_1.3.0
[10] glue_1.3.0 withr_2.1.2 R.utils_2.7.0
[13] modelr_0.1.2 readxl_1.1.0 bindr_0.1.1
[16] plyr_1.8.4 munsell_0.5.0 gtable_0.2.0
[19] cellranger_1.1.0 rvest_0.3.2 R.methodsS3_1.7.1
[22] evaluate_0.11 labeling_0.3 knitr_1.20
[25] broom_0.5.0 Rcpp_0.12.19 scales_1.0.0
[28] backports_1.1.2 jsonlite_1.5 hms_0.4.2
[31] digest_0.6.17 stringi_1.2.4 grid_3.5.1
[34] rprojroot_1.3-2 cli_1.0.1 tools_3.5.1
[37] magrittr_1.5 lazyeval_0.2.1 crayon_1.3.4
[40] whisker_0.3-2 pkgconfig_2.0.2 xml2_1.2.0
[43] lubridate_1.7.4 assertthat_0.2.0 rmarkdown_1.10
[46] httr_1.3.1 rstudioapi_0.8 R6_2.3.0
[49] nlme_3.1-137 git2r_0.23.0 compiler_3.5.1
This reproducible R Markdown analysis was created with workflowr 1.1.1