Last updated: 2023-08-30

Checks: 6 1

Knit directory: duplex_sequencing_screen/

This reproducible R Markdown analysis was created with workflowr (version 1.6.2). The Checks tab describes the reproducibility checks that were applied when the results were created. The Past versions tab lists the development history.


The R Markdown file has unstaged changes. To know which version of the R Markdown file created these results, you’ll want to first commit it to the Git repo. If you’re still working on the analysis, you can ignore this warning. When you’re finished, you can run wflow_publish to commit the R Markdown file and build the HTML.

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.

The command set.seed(20200402) 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.

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Nice! There were no cached chunks for this analysis, so you can be confident that you successfully produced the results during this run.

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Great! You are using Git for version control. Tracking code development and connecting the code version to the results is critical for reproducibility.

The results in this page were generated with repository version f9eea37. See the Past versions tab to see a history of the changes made to the R Markdown and HTML files.

Note that you need to be careful to ensure that all relevant files for the analysis have been committed to Git prior to generating the results (you can use 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:


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Unstaged changes:
    Modified:   .DS_Store
    Modified:   analysis/ABL_kinasefunction_data_parser.Rmd
    Modified:   code/compare_screens.R
    Modified:   data/Consensus_Data/novogene_lane18/tlane18a_sample6/.DS_Store
    Modified:   output/ABLEnrichmentScreens/ABL_Region1_Lane18_Comparisons/.DS_Store
    Modified:   output/ABLEnrichmentScreens/ABL_Region1_Lane18_Comparisons/cross-replicate/.DS_Store

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.


These are the previous versions of the repository in which changes were made to the R Markdown (analysis/ABL_kinasefunction_data_parser.Rmd) and HTML (docs/ABL_kinasefunction_data_parser.html) files. If you’ve configured a remote Git repository (see ?wflow_git_remote), click on the hyperlinks in the table below to view the files as they were in that past version.

File Version Author Date Message
Rmd f9eea37 haiderinam 2023-08-30 2023 Updates

#Cleanup code for plotting
source("code/plotting/cleanup.R")

#8.30.23 #The following code was taken from ABL_SM_CRISPR_Cut_Analyses.Rmd #Essentially all this code does is that it takes two BaF3 ABL Region 1 D0 samples and two D6 samples, and creates a dataframe with enrichment scores. #Ivan wanted counts from separate D0 backgrounds, and he wanted every mutant, regardless of whether it was seen in both baseline samples (when I do analyses, I take out any mutant that is not seen in both baseline samples). Let’s call these “unseen mutants” #This code does not include unseen mutants by default. To make sure that unseen mutants are seen, uncomment out the Ivan section in compare_screens.R. Once you uncomment it, then run the code below #One more note: this is Region 1 (Amino acids 242-322) data from Lane 18 only. In the future you can include samples from other lanes.

source("code/compare_screens.R")

source("code/plotting/cleanup.R")
source("code/plotting/heatmap_plotting_function.R")
# rm(list=ls())

comparisons=read.csv("data/Consensus_Data/novogene_lane18/TwistRegion1Screen_Comparisons_Todo.csv")
# comparisons=comparisons%>%filter(Completed%in%"FALSE")

# for(i in 1:nrow(comparisons)){
  i=10
  dirname=comparisons$dirname[i]
  pathname=paste("output/ABLEnrichmentScreens/ABL_Region1_Lane18_Comparisons/",dirname,sep = "")
  # Create directory if it doesn't already exist
  if (!file.exists(pathname)){
    dir.create(pathname)
} 
before_screen1_identifier=unlist(strsplit(comparisons$before_screen1_identifier[i],","))
after_screen1_identifier=unlist(strsplit(comparisons$after_screen1_identifier[i],","))
before_screen2_identifier=unlist(strsplit(comparisons$before_screen2_identifier[i],","))
before_screen2_identifier="BTRID0_FT"
after_screen2_identifier=unlist(strsplit(comparisons$after_screen2_identifier[i],","))
  # length(after_screen1_identifier)
# screen_compare_means=compare_screens(comparisons$before_screen1_identifier[i],
#                                      comparisons$after_screen1_identifier[i],
#                                      comparisons$before_screen2_identifier[i],
#                                      comparisons$after_screen2_identifier[i])
screen_compare_means=compare_screens(before_screen1_identifier,
                                     after_screen1_identifier,
                                     before_screen2_identifier,
                                     after_screen2_identifier)

screen_compare_means_forexport=apply(screen_compare_means,2,as.character)
# dirname
# write.csv(screen_compare_means_forexport,file="output/ABLEnrichmentScreens/ABL_Region1_Lane18_Comparisons/cross-replicate/baf3_IL3_rep1vsrep2_ft/screen_comparison_baf3_IL3_low_rep1vsrep2ft.csv")

# Plot 1. What does the heatmap look like from the average of the net growth rate?
heatmap_plotting_function(screen_compare_means,242,321,fill_variable = "netgr_obs_mean",fill_name = "Net growth rate")

# ggsave(paste("output/ABLEnrichmentScreens/ABL_Region1_Lane18_Comparisons/",dirname,"/plot1_heatmap.pdf",sep=""),width=10,height=6,units="in",useDingbats=F)
# screen_compare_means2=screen_compare_means%>%filter(alt_codon%in%twist$Codon)

# Plot 2a: What do the correlations look like for net growth rate (show mutants in text)?

ggplot(screen_compare_means,aes(x=netgr_obs_screen1,y=netgr_obs_screen2,color=resmuts,label=species))+geom_text(size=2.5)+geom_abline()+cleanup+stat_cor(method = "pearson")+labs(color="Known\nResistant\nMutant")+scale_x_continuous("Net growth rate screen 1")+scale_y_continuous("Net growth rate screen 2")

# ggsave(paste("output/ABLEnrichmentScreens/ABL_Region1_Lane18_Comparisons/",dirname,"/plot2a_Netgrowthrate_correlations_text.pdf",sep=""),width=6,height=4,units="in",useDingbats=F)

# Plot 2b: What do the correlations look like for enrichment scores (show mutants in points)?
ggplot(screen_compare_means,aes(x=netgr_obs_screen1,y=netgr_obs_screen2,label=species))+geom_point(color="black",shape=21,size=2,aes(fill=resmuts))+geom_abline()+cleanup+stat_cor(method = "pearson")+labs(fill="Known\nResistant\nMutant")+scale_x_continuous("Net growth rate screen 1")+scale_y_continuous("Net growth rate screen 2")

# ggsave(paste("output/ABLEnrichmentScreens/ABL_Region1_Lane18_Comparisons/",dirname,"/plot2b_Netgrowthrate_correlations_points.pdf",sep=""),width=6,height=4,units="in",useDingbats=F)
# Plot 2c: What do the correlations look like for enrichment scores (show mutants in text)?
ggplot(screen_compare_means,aes(x=score_screen1,y=score_screen2,color=resmuts,label=species))+geom_text(size=2.5)+geom_abline()+cleanup+ stat_cor(method = "pearson")+labs(color="Known\nResistant\nMutant")+scale_x_continuous("Enrichment score screen 1")+scale_y_continuous("Enrichment score screen 2")

# ggsave(paste("output/ABLEnrichmentScreens/ABL_Region1_Lane18_Comparisons/",dirname,"/plot2c_Enrichmentscores_correlations_text.pdf",sep=""),width=6,height=4,units="in",useDingbats=F)

# Plot 3a: Plots: what are the overall net growth rate distributions?
ggplot(screen_compare_means,aes(x=netgr_obs_mean,fill=resmuts))+geom_density(alpha=0.7)+cleanup+labs(fill="Known\nResistant\nMutant")+scale_x_continuous("Mean net growth rate of screens")

# ggsave(paste("output/ABLEnrichmentScreens/ABL_Region1_Lane18_Comparisons/",dirname,"/plot3a_Netgrowthrate_distributions_resmuts.pdf",sep=""),width=6,height=4,units="in",useDingbats=F)
# Plot 3b: Plots: what are the net growth rate distributions?

library(reshape2)
screen_compare_melt=melt(screen_compare_means%>%dplyr::select(species,netgr_obs_screen1,netgr_obs_screen2),id.vars = "species",measure.vars =c("netgr_obs_screen1","netgr_obs_screen2"),variable.name = "Condition",value.name = "netgr_obs")


ggplot(screen_compare_melt,aes(x=netgr_obs,fill=Condition))+
  geom_density(alpha=0.7)+
  cleanup+
  scale_x_continuous("Net growth rate observed")+
  scale_fill_discrete(labels=c("Screen 1","Screen 2"))

# ggsave(paste("output/ABLEnrichmentScreens/ABL_Region1_Lane18_Comparisons/",dirname,"/plot3b_Netgrowthrate_distributions.pdf",sep=""),width=6,height=4,units="in",useDingbats=F)

# }

# dirname="K562_Medium_rep1vs2"

sessionInfo()
R version 4.0.0 (2020-04-24)
Platform: x86_64-apple-darwin17.0 (64-bit)
Running under: macOS  10.16

Matrix products: default
BLAS:   /Library/Frameworks/R.framework/Versions/4.0/Resources/lib/libRblas.dylib
LAPACK: /Library/Frameworks/R.framework/Versions/4.0/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  stats     graphics  grDevices utils     datasets  methods  
[8] base     

other attached packages:
 [1] reshape2_1.4.4     ggpubr_0.4.0       RColorBrewer_1.1-2 doParallel_1.0.15 
 [5] iterators_1.0.12   foreach_1.5.0      tictoc_1.0         plotly_4.9.2.1    
 [9] ggplot2_3.3.3      dplyr_1.0.6        stringr_1.4.0     

loaded via a namespace (and not attached):
 [1] httr_1.4.2        sass_0.4.1        tidyr_1.1.3       jsonlite_1.7.2   
 [5] viridisLite_0.3.0 carData_3.0-3     bslib_0.3.1       assertthat_0.2.1 
 [9] cellranger_1.1.0  yaml_2.2.1        pillar_1.6.1      backports_1.1.7  
[13] glue_1.4.1        digest_0.6.25     promises_1.1.0    ggsignif_0.6.0   
[17] colorspace_1.4-1  htmltools_0.5.2   httpuv_1.5.2      plyr_1.8.6       
[21] pkgconfig_2.0.3   broom_0.7.6       haven_2.4.1       purrr_0.3.4      
[25] scales_1.1.1      whisker_0.4       openxlsx_4.1.5    later_1.0.0      
[29] rio_0.5.16        git2r_0.27.1      tibble_3.1.2      generics_0.0.2   
[33] farver_2.0.3      car_3.0-7         ellipsis_0.3.2    withr_2.4.2      
[37] lazyeval_0.2.2    magrittr_2.0.1    crayon_1.4.1      readxl_1.3.1     
[41] evaluate_0.14     fs_1.4.1          fansi_0.4.1       rstatix_0.6.0    
[45] forcats_0.5.1     foreign_0.8-78    tools_4.0.0       data.table_1.12.8
[49] hms_1.1.0         lifecycle_1.0.0   munsell_0.5.0     zip_2.0.4        
[53] compiler_4.0.0    jquerylib_0.1.4   rlang_0.4.11      grid_4.0.0       
[57] htmlwidgets_1.5.1 labeling_0.3      rmarkdown_2.14    gtable_0.3.0     
[61] codetools_0.2-16  abind_1.4-5       DBI_1.1.0         curl_4.3         
[65] R6_2.4.1          knitr_1.28        fastmap_1.1.0     utf8_1.1.4       
[69] workflowr_1.6.2   rprojroot_1.3-2   stringi_1.7.5     Rcpp_1.0.4.6     
[73] vctrs_0.3.8       tidyselect_1.1.0  xfun_0.31