Differential RNA splicing analysis using leafcutter¶
Summary¶
Differential splicing analysis using leafcutter, one of the best tools.
Only works for hg19 or hg38.
Only works for paired-end RNA-seq data.
Input¶
Example data dir: Projects/Siqi_data/RNAseq_data/Differential_Exon_Analysis/test
1. fastq.tsv¶
3-col tsv, R1 R2 label.
Use run_lsf.py --guess_input to generate.
2. design.tsv¶
Two columns. First column should match to the labels in fastq.tsv. Second col is the group. Only pairwise. From leafcutter: Additional columns can be used to specify confounders, e.g. batch/sex/age. Numeric columns will be treated as continuous, so use e.g. batch1, batch2, batch3 rather than 1, 2, 3 if you a categorical variable.
2393110_Nontarget_Hudep2_2 WT
2393109_Nontarget_Hudep2_1 WT
2393111_Nontarget_Hudep2 WT
2393114_M1_Hudep2 sgRNA
2393113_M1_2_Hudep2_2 sgRNA
2393112_M1_2_Hudep2_1 sgRNA
Usage¶
hpcf_interactive
module load python/2.7.13
run_lsf.py -f fastq.tsv -p leafcutter -g hg38 --design_matrix $PWD/design.tsv
Output¶
Results are stored in the $jid folder.