renamed files to make more generic
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meta_data_analysis/AF_and_OR_calcs.R
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meta_data_analysis/AF_and_OR_calcs.R
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#============================================
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# TASK: To calculate Allele Frequency and
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# Odds Ratio from master data
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# and add the calculated params to meta_data extracted from
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# pnca_data_extraction.py
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#===========================================
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homedir = '~'
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getwd()
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#setwd('~/git/LSHTM_analysis/meta_data_analysis')
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setwd(paste0(homedir, '/', 'git/LSHTM_analysis/meta_data_analysis'))
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getwd()
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#%% variable assignment: input and output paths & filenames
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drug = 'pyrazinamide'
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gene = 'pncA'
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gene_match = paste0(gene,'_p.')
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print(gene_match)
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#=======
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# input dir
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#=======
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# file1: Raw data
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#indir = 'git/Data/pyrazinamide/input/original'
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indir = paste0('git/Data', '/', drug, '/', 'input/original')
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in_filename = 'original_tanushree_data_v2.csv'
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infile = paste0(homedir, '/', indir, '/', in_filename)
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print(paste0('Reading infile:', ' ', infile) )
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# file2: file to extract valid snps and add calcs to: pnca_metadata.csv {outfile3 from data extraction script}
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indir_metadata = paste0('git/Data', '/', drug, '/', 'output')
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in_filename_metadata = 'pnca_metadata.csv'
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infile_metadata = paste0(homedir, '/', indir_metadata, '/', in_filename_metadata)
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print(paste0('Reading metadata infile:', infile_metadata))
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#=========
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# output dir
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#=========
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# output filename in respective section at the time of outputting files
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#outdir = 'git/Data/pyrazinamide/output'
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outdir = paste0('git/Data', '/', drug, '/', 'output')
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out_filename = paste0(tolower(gene),'_', 'meta_data_with_AFandOR.csv')
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outfile = paste0(homedir, '/', outdir, '/', out_filename)
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print(paste0('Output file with full path:', outfile))
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#%% end of variable assignment for input and output files
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#===============
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# Step 1: Read master/raw data stored in Data/
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#===============
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raw_data_all = read.csv(infile, stringsAsFactors = F)
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raw_data = raw_data_all[,c("id"
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, "pyrazinamide"
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, "dr_mutations_pyrazinamide"
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, "other_mutations_pyrazinamide")]
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rm(raw_data_all)
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rm(indir, in_filename, infile)
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#####
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# 1a: exclude na
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#####
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raw_data = raw_data[!is.na(raw_data$pyrazinamide),]
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total_samples = length(unique(raw_data$id))
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print(paste0('Total samples without NA in', ' ', drug, 'is:', total_samples))
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# sanity check: should be true
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is.numeric(total_samples)
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#####
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# 1b: combine the two mutation columns
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#####
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raw_data$all_mutations_pyrazinamide = paste(raw_data$dr_mutations_pyrazinamide
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, raw_data$other_mutations_pyrazinamide)
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head(raw_data$all_mutations_pyrazinamide)
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#####
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# 1c: create yet another column that contains all the mutations but in lower case
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#####
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raw_data$all_muts_pnca = tolower(raw_data$all_mutations_pyrazinamide)
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# sanity checks
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#table(grepl("pnca_p",raw_data$all_muts_pnca))
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print(paste0('converting gene match:', gene_match, ' ', 'to lowercase'))
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gene_match = tolower(gene_match)
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table(grepl(gene_match,raw_data$all_muts_pnca))
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# sanity check: should be TRUE
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#sum(table(grepl("pnca_p",raw_data$all_muts_pnca))) == total_samples
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sum(table(grepl(gene_match,raw_data$all_muts_pnca))) == total_samples
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# set up variables: can be used for logistic regression as well
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i = "pnca_p.ala134gly" # has a NA, should NOT exist
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table(grepl(i,raw_data$all_muts_pnca))
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i = "pnca_p.trp68gly"
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table(grepl(i,raw_data$all_muts_pnca))
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mut = grepl(i,raw_data$all_muts_pnca)
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dst = raw_data$pyrazinamide
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table(mut, dst)
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#chisq.test(table(mut,dst))
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#fisher.test(table(mut, dst))
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#table(mut)
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#===============
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# Step 2: Read valid snps for which OR can be calculated (infile_comp_snps.csv)
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#===============
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print(paste0('Reading metadata infile:', infile_metadata))
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pnca_metadata = read.csv(infile_metadata
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# , file.choose()
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, stringsAsFactors = F
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, header = T)
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# clear variables
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rm(homedir, in_filename, indir, infile)
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rm(indir_metadata, infile_metadata, in_filename_metadata)
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# count na in pyrazinamide column
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tot_pza_na = sum(is.na(pnca_metadata$pyrazinamide))
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expected_rows = nrow(pnca_metadata) - tot_pza_na
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# drop na from the pyrazinamide colum
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pnca_snps_or = pnca_metadata[!is.na(pnca_metadata$pyrazinamide),]
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# sanity check
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if(nrow(pnca_snps_or) == expected_rows){
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print('PASS: no. of rows match with expected_rows')
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} else{
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print('FAIL: nrows mismatch.')
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}
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# extract unique snps to iterate over for AF and OR calcs
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pnca_snps_unique = unique(pnca_snps_or$mutation)
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print(paste0('Total no. of distinct comp snps to perform OR calcs: ', length(pnca_snps_unique)))
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# Define OR function
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x = as.numeric(mut)
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y = dst
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or = function(x,y){
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tab = as.matrix(table(x,y))
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a = tab[2,2]
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if (a==0){ a<-0.5}
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b = tab[2,1]
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if (b==0){ b<-0.5}
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c = tab[1,2]
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if (c==0){ c<-0.5}
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d = tab[1,1]
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if (d==0){ d<-0.5}
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(a/b)/(c/d)
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}
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dst = raw_data$pyrazinamide
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ors = sapply(pnca_snps_unique,function(m){
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mut = grepl(m,raw_data$all_muts_pnca)
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or(mut,dst)
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})
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ors
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pvals = sapply(pnca_snps_unique,function(m){
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mut = grepl(m,raw_data$all_muts_pnca)
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fisher.test(mut,dst)$p.value
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})
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pvals
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afs = sapply(pnca_snps_unique,function(m){
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mut = grepl(m,raw_data$all_muts_pnca)
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mean(mut)
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})
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afs
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# check ..hmmm
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afs['pnca_p.trp68gly']
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afs['pnca_p.gln10pro']
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afs['pnca_p.leu4ser']
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plot(density(log(ors)))
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plot(-log10(pvals))
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hist(log(ors)
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, breaks = 100
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)
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# FIXME: could be good to add a sanity check
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if (table(names(ors) == names(pvals)) & table(names(ors) == names(afs)) & table(names(pvals) == names(afs)) == length(pnca_snps_unique)){
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print('PASS: names of ors, pvals and afs match: proceed with combining into a single df')
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} else{
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print('FAIL: names of ors, pvals and afs mismatch')
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}
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# combine
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comb_AF_and_OR = data.frame(ors, pvals, afs)
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head(rownames(comb_AF_and_OR))
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# add rownames of comb_AF_and_OR as an extra column 'mutation' to allow merging based on this column
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comb_AF_and_OR$mutation = rownames(comb_AF_and_OR)
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# sanity check
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if (table(rownames(comb_AF_and_OR) == comb_AF_and_OR$mutation)){
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print('PASS: rownames and mutaion col values match')
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}else{
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print('FAIL: rownames and mutation col values mismatch')
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}
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############
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# Merge 1:
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###########
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df1 = pnca_metadata
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df2 = comb_AF_and_OR
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head(df1$mutation); head(df2$mutation)
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# FIXME: newlines
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print(paste0('merging two dfs: '
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,'\ndf1 (big df i.e. meta data) nrows: ', nrow(df1)
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,'\ndf2 (small df i.e af, or, pval) nrows: ', nrow(df2)
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, 'expected rows in merged df: ', nrow(df1), 'expected cols in merged_df: ', (ncol(df1) + ncol(df2) - 1)))
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merged_df = merge(df1 # big file
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, df2 # small (afor file)
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, by = "mutation"
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, all.x = T) # because you want all the entries of the meta data
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# sanity check
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if(ncol(merged_df) == (ncol(df1) + ncol(df2) - 1)){
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print(paste0('PASS: no. of cols is as expected: ', ncol(merged_df)))
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} else{
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print('FAIL: no.of cols mistmatch')
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}
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# quick check
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i = "pnca_p.ala134gly" # has all NAs in pyrazinamide, should be NA in ors, etc.
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merged_df[merged_df$mutation == i,]
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# count na in each column
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na_count = sapply(merged_df, function(y) sum(length(which(is.na(y))))); na_count
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# only some or and Af should be NA
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#Row.names ors pvals afs
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#63 63 63 63
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# reassign custom colnames
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colnames(merged_df)[colnames(merged_df)== "ors"] <- "OR"
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colnames(merged_df)[colnames(merged_df)== "afs"] <- "AF"
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colnames(merged_df)[colnames(merged_df)== "pvals"] <- "pvalue"
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colnames(merged_df)
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# add log OR and neglog pvalue
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merged_df$logor = log(merged_df$OR)
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is.numeric(merged_df$logor)
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merged_df$neglog10pvalue = -log10(merged_df$pvalue)
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is.numeric(merged_df$neglog10pvalue)
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merged_df$AF_percent = merged_df$AF*100
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is.numeric(merged_df$AF_percent)
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# check AFs
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#i = 'pnca_p.trp68gly'
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i = 'pnca_p.gln10pro'
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#i = 'pnca_p.leu4ser'
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merged_df[merged_df$mutation == i,]
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# FIXME: harcoding (beware!), NOT FATAL though!
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ncol_added = 3
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print(paste0('Added', ncol_added, ' ', 'more cols to merged_df i.e log10 OR and -log10 P-val: '
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, 'no. of cols in merged_df now: ', ncol(merged_df)))
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#%% write file out: pnca_meta_data_with_AFandOR
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print(paste0('writing output file in: '
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, 'Filename: ', out_filename
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, 'Path:', outdir))
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write.csv(merged_df, outfile
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, row.names = F)
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print(paste0('Finished writing:', out_filename, '\nExpected no. of cols:', ncol(merged_df)))
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print('======================================================================')
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rm(out_filename)
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