All Perl, Python, R, and Bash Shell scripts were used to eQTLs and RNA-m6A QTLs analysis by integrating QTLtools and MASH.
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Quantify reads density from ChIP-seq or MeRIP-seq data by
1a_QTLtools_quan.peaks.pl, or quantify gene expression level from RNA-seq data by1b_QTLtools_quan.gene.pl.QTLtools quanis invoked in line 125, GTF file may need to be changed based on your file name. More details please seeperl 1a_QTLtools_quan.peaks.pl -helporperl 1b_QTLtools_quan.gene.pl -help. -
Get value log2((FPKM_IP+1)/(FPKM_INPUT+1)) of each peak for each MeRIP-seq sample by
2a_log2ratio.peaks.pl. And get value log2(TPM_INPUT+1) of each gene for each RNA-seq sample (Input of MeRIP-seq) by2b_log2TPM.gene.pl. More details please seeperl xxx.pl -help. -
Merge all files from each sample into one file to satisfy the input format of
QTLtools cisandQTLtools transby3a_merge.peaks.plor3b_merge.gene.pl. Please ensure that the colomn orders of the GFT file and the merged file are the same. More details please seeperl xxx.pl -help. -
Further filter m6A-epaks or genes by
4a_keptRegions.peaks.Ror4b_keptRegions.gene.R. We can remove peaks or genes with low reads desnity or low variance. More details please see code of4a_keptRegions.peaks.Ror4b_keptRegions.gene.R -
PCA for genotype by
5_QTLtools_pca.SNP.sh. PCA and PEER factor analysis for phenotype please see the codes under foldercovariates.
6-8. Identify cis- or trans-QTLs by invoking QTLtools cis or QTLtools trans.