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Overview

  • Similar to exercise 6.4 we will:

    • Use created “samplesheet.csv” metadata file for small RNAseq datasets in exercise 6.4.

    • Use a “nextflow.config” file in the working directory to override Nextflow parameters (e.g., specify where to find the pipeline assets).

    • Use a PBS script to run the expression profiling of miRNAs against MirGeneDB, a curated database that includes experimentally validated miRNAs.

Prepare pipeline inputs

Let’s move to the working directory:

Code Block
cd $HOME/workshop/2024-2/session6_smallRNAseq/runs/run2_human_MirGeneDB

Now, let’s copy the samplesheet.csv, nextflow.config and the PBS script to run the pipeline agains MirGeneDB files:

Code Block
cp $HOME/workshop/2024-2/session6_smallRNAseq/data/human_disease/samplesheet.csv .
cp $HOME/workshop/2024-2/session6_smallRNAseq/scripts/nextflow.config .
cp $HOME/workshop/2024-2/session6_smallRNAseq/scripts/launch_nf-core_smallRNAseq_MirGeneDB.pbs .

Print the content of the launch script:

Code Block
cat launch_nf-core_smallRNAseq_MirGeneDB.pbs

...

Submit the job to the HPC cluster:

Code Block
qsub launch_nf-core_smallRNAseq_MirGeneDB.pbs

Monitor the progress:

Code Block
qjobs

The job will take several hours to run, hence we will use precomputed results for the statistical analysis in the next section.

Outputs

The pipeline will produce two folders, one called “work,” where all the processing is done, and another called “results,” where we can find the pipeline's outputs. The content of the results folder is as follows:

Code Block
results/
├── bowtie_index
│   ├── mirna_hairpin
│   └── mirna_mature
├── fastp
│   └── on_raw
├── fastqc
│   ├── raw
│   └── trimmed
├── mirna_quant
│   ├── bam
│   ├── edger_qc <-- Expression mature miRNA (mature_counts.csv) and precursor-miRNAs (haripin_counts.csv)
│   ├── mirtop
│   ├── reference
│   └── seqcluster
├── mirtrace
│   ├── mirtrace-report.html
│   ├── mirtrace-results.json
│   ├── mirtrace-stats-contamination_basic.tsv
│   ├── mirtrace-stats-contamination_detailed.tsv
│   ├── mirtrace-stats-length.tsv
│   ├── mirtrace-stats-mirna-complexity.tsv
│   ├── mirtrace-stats-phred.tsv
│   ├── mirtrace-stats-qcstatus.tsv
│   ├── mirtrace-stats-rnatype.tsv
│   ├── qc_passed_reads.all.collapsed
│   └── qc_passed_reads.rnatype_unknown.collapsed
├── multiqc
│   ├── multiqc_data
│   ├── multiqc_plots
│   └── multiqc_report.html
└── pipeline_info
    ├── execution_report_2024-08-26_14-38-10.html
    ├── execution_report_2024-08-26_17-44-50.html
    ├── execution_timeline_2024-08-26_14-38-10.html
    ├── execution_timeline_2024-08-26_17-44-50.html
    ├── execution_trace_2024-08-26_17-44-50.txt
    ├── nf_core_smrnaseq_software_mqc_versions.yml
    ├── params_2024-08-26_17-45-00.json
    ├── pipeline_dag_2024-08-26_14-38-10.html
    └── pipeline_dag_2024-08-26_17-44-50.html

The quantification of the mature miRNA and hairpin expressions can be found in the /results/mirna_quant/edger_qc directory.

Code Block
cd /results/mirna_quant/edger_qc
Code Block
├── hairpin_counts.csv
├── hairpin_CPM_heatmap.pdf
├── hairpin_edgeR_MDS_distance_matrix.txt
├── hairpin_edgeR_MDS_plot_coordinates.txt
├── hairpin_edgeR_MDS_plot.pdf
├── hairpin_log2CPM_sample_distances_dendrogram.pdf
├── hairpin_log2CPM_sample_distances_heatmap.pdf
├── hairpin_log2CPM_sample_distances.txt
├── hairpin_logtpm.csv
├── hairpin_logtpm.txt
├── hairpin_normalized_CPM.txt
├── hairpin_unmapped_read_counts.txt
├── mature_counts.csv  <-- Expression profile of mature miRNAs.
├── mature_CPM_heatmap.pdf
├── mature_edgeR_MDS_distance_matrix.txt
├── mature_edgeR_MDS_plot_coordinates.txt
├── mature_edgeR_MDS_plot.pdf
├── mature_log2CPM_sample_distances_dendrogram.pdf
├── mature_log2CPM_sample_distances_heatmap.pdf
├── mature_log2CPM_sample_distances.txt
├── mature_logtpm.csv
├── mature_logtpm.txt
├── mature_normalized_CPM.txt
└── mature_unmapped_read_counts.txt

Let’s inspect the mature.csv file. Let’s use the ‘cat’ command to print it on the screen:

Code Block
cat mature_counts.csv
Code Block
"","Hsa-Let-7-P1b_5p","Hsa-Let-7-P1c_5p","Hsa-Let-7-P1d_5p","Hsa-Let-7-P2a1_5p","Hsa-Let-7-P2a2_5p","Hsa-Let-7-P2a3_5p","Hsa-Let-7-P2b1_5p","Hsa-Let-7-P2b2_5p","Hsa-Let-7-P2b3_5p","Hsa-Let-7-P2c1_5p","Hsa-Let-7-P2c2_5p","Hsa-Let-7-P2c3_5p","Hsa-Mir-1-P1_3p","Hsa-Mir-1-P2_3p","Hsa-Mir-1-P3_3p","Hsa-Mir-7-P1_5p","Hsa-Mir-7-P2_5p","Hsa-Mir-7-P4_5p","Hsa-Mir-8-P1a_3p","Hsa-Mir-8-P1b_3p","Hsa-Mir-8-P2a_3p","Hsa-Mir-8-P2b_3p","Hsa-Mir-8-P3a_3p","Hsa-Mir-9-P1_5p","Hsa-Mir-9-P1_3p","Hsa-Mir-9-P2_5p","Hsa-Mir-9-P2_3p","Hsa-Mir-9-P3_5p","Hsa-Mir-9-P3_3p","Hsa-Mir-10-P1b-v1_5p","Hsa-Mir-10-P1b-v2_5p","Hsa-Mir-10-P1c-v1_5p","Hsa-Mir-10-P1c-v2_5p","Hsa-Mir-10-P2b_5p","Hsa-Mir-10-P2c_5p","Hsa-Mir-10-P2d_5p","Hsa-Mir-10-P3b_5p","Hsa-Mir-10-P3c_5p","Hsa-Mir-10-P3d_5p","Hsa-Mir-15-P1a_5p","Hsa-Mir-15-P1b_5p","Hsa-Mir-15-P1c_5p","Hsa-Mir-15-P1d_5p","Hsa-Mir-15-P2a_5p","Hsa-Mir-15-P2b_5p","Hsa-Mir-15-P2c_5p","Hsa-Mir-15-P2d_5p","Hsa-Mir-17-P1a_5p","Hsa-Mir-17-P1c_5p","Hsa-Mir-17-P1d_5p","Hsa-Mir-17-P1d_3p","Hsa-Mir-17-P2a_5p","Hsa-Mir-17-P2c_5p","Hsa-Mir-17-P4a_5p","Hsa-Mir-17-P4c_5p","Hsa-Mir-17-P4d_5p","Hsa-Mir-19-P1a_3p","Hsa-Mir-19-P2a_3p","Hsa-Mir-19-P2c_3p","Hsa-Mir-21_5p","Hsa-Mir-22-P1b_3p","Hsa-Mir-23-P2_3p","Hsa-Mir-23-P3_3p","Hsa-Mir-24-P2_3p","Hsa-Mir-24-P3_3p","Hsa-Mir-26-P1_5p","Hsa-Mir-26-P2_5p","Hsa-Mir-26-P4_5p","Hsa-Mir-27-P2_3p","Hsa-Mir-27-P3_3p","Hsa-Mir-28-P1_5p","Hsa-Mir-28-P1_3p","Hsa-Mir-28-P2_5p","Hsa-Mir-28-P2_3p","Hsa-Mir-28-P3_5p","Hsa-Mir-29-P1b-v1_3p","Hsa-Mir-29-P1b-v2_3p","Hsa-Mir-29-P1d-v1_3p","Hsa-Mir-29-P1d-v2_3p","Hsa-Mir-29-P2b_3p","Hsa-Mir-29-P2d_3p","Hsa-Mir-30-P1a_5p","Hsa-Mir-30-P1b_5p","Hsa-Mir-30-P1d_5p","Hsa-Mir-30-P2a_5p","Hsa-Mir-30-P2b_5p","Hsa-Mir-30-P2d_5p","Hsa-Mir-31_5p","Hsa-Mir-32_5p","Hsa-Mir-33-P1_5p","Hsa-Mir-33-P3_5p","Hsa-Mir-34-P1_5p","Hsa-Mir-34-P2a_5p","Hsa-Mir-34-P2a_3p","Hsa-Mir-34-P2b_5p","Hsa-Mir-34-P3a-v1_5p","Hsa-Mir-34-P3a-v2_5p","Hsa-Mir-34-P3b_5p","Hsa-Mir-34-P3b_3p","Hsa-Mir-34-P3c_5p","Hsa-Mir-92-P1a_3p","Hsa-Mir-92-P1c_3p","Hsa-Mir-92-P1d_3p","Hsa-Mir-92-P2a_3p","Hsa-Mir-92-P2c_3p","Hsa-Mir-92-P2d_3p","Hsa-Mir-95-P1_3p","Hsa-Mir-95-P2_3p","Hsa-Mir-95-P3-v1_5p","Hsa-Mir-95-P3-v2_5p","Hsa-Mir-95-P4_5p","Hsa-Mir-95-P4_3p","Hsa-Mir-96-P1_5p","Hsa-Mir-96-P2_5p","Hsa-Mir-96-P3-v1_5p","Hsa-Mir-96-P3-v2_5p","Hsa-Mir-101-P1-v1_3p","Hsa-Mir-101-P1-v2_3p","Hsa-Mir-101-P2-v1_3p","Hsa-Mir-101-P2-v2_3p","Hsa-Mir-103-P1_3p","Hsa-Mir-103-P2_3p","Hsa-Mir-103-P4_3p","Hsa-Mir-105-P1_5p","Hsa-Mir-105-P2_5p","Hsa-Mir-122_5p","Hsa-Mir-124-P1-v1_3p","Hsa-Mir-124-P1-v2_3p","Hsa-Mir-124-P2-v1_3p","Hsa-Mir-124-P2-v2_3p","Hsa-Mir-124-P3-v1_3p","Hsa-Mir-124-P3-v2_3p","Hsa-Mir-126-P2-v1_3p","Hsa-Mir-126-P2-v2_5p","Hsa-Mir-126-P2-v3_3p","Hsa-Mir-127_3p","Hsa-Mir-128-P1_3p","Hsa-Mir-128-P2_3p","Hsa-Mir-129-P1_3p","Hsa-Mir-129-P2_5p","Hsa-Mir-129-P2_3p","Hsa-Mir-130-P1c_3p","Hsa-Mir-130-P2a_3p","Hsa-Mir-130-P2b_3p","Hsa-Mir-130-P3b_3p","Hsa-Mir-130-P4a_5p","Hsa-Mir-130-P4a_3p","Hsa-Mir-132-P1_3p","Hsa-Mir-132-P2_5p","Hsa-Mir-132-P2_3p","Hsa-Mir-133-P1-v1_3p","Hsa-Mir-133-P1-v2_3p","Hsa-Mir-133-P2-v1_3p","Hsa-Mir-133-P2-v2_3p","Hsa-Mir-133-P3-v1_3p","Hsa-Mir-133-P3-v2_3p","Hsa-Mir-134_5p","Hsa-Mir-135-P1_5p","Hsa-Mir-135-P2_5p","Hsa-Mir-135-P4_5p","Hsa-Mir-136-v1_3p","Hsa-Mir-136-v2_5p","Hsa-Mir-136-v2_3p","Hsa-Mir-137-P1-v1_3p","Hsa-Mir-137-P1-v2_3p","Hsa-Mir-138-P1_5p","Hsa-Mir-138-P2_5p","Hsa-Mir-139_5p","Hsa-Mir-140-P1-v1_3p","Hsa-Mir-140-P1-v2_3p","Hsa-Mir-142-P1-v1_5p","Hsa-Mir-142-P1-v2_3p","Hsa-Mir-142-P1-v3_3p","Hsa-Mir-142-P1-v4_5p","Hsa-Mir-143_3p","Hsa-Mir-144-v1_3p","Hsa-Mir-144-v2_3p","Hsa-Mir-145_5p","Hsa-Mir-146-P1_5p","Hsa-Mir-146-P4_5p","Hsa-Mir-147-v1_3p","Hsa-Mir-147-v2_3p","Hsa-Mir-148-P1_3p","Hsa-Mir-148-P3_3p","Hsa-Mir-148-P4_3p","Hsa-Mir-149_5p","Hsa-Mir-150_5p","Hsa-Mir-153-P1_3p","Hsa-Mir-153-P2_3p","Hsa-Mir-154-P1_5p","Hsa-Mir-154-P1_3p","Hsa-Mir-154-P2-v1_5p","Hsa-Mir-154-P2-v2_5p","Hsa-Mir-154-P3a-v1_3p","Hsa-Mir-154-P3a-v2_3p","Hsa-Mir-154-P3b_3p","Hsa-Mir-154-P4a_3p","Hsa-Mir-154-P4b_3p","Hsa-Mir-154-P5_5p","Hsa-Mir-154-P5_3p","Hsa-Mir-154-P6_3p","Hsa-Mir-154-P7_5p","Hsa-Mir-154-P8_5p","Hsa-Mir-154-P8_3p","Hsa-Mir-154-P9_3p","Hsa-Mir-154-P10_5p","Hsa-Mir-154-P12_3p","Hsa-Mir-154-P13-v1_5p","Hsa-Mir-154-P13-v2_5p","Hsa-Mir-154-P14_5p","Hsa-Mir-154-P15_5p","Hsa-Mir-154-P16_3p","Hsa-Mir-154-P17_3p","Hsa-Mir-154-P18_3p","Hsa-Mir-154-P19_3p","Hsa-Mir-154-P20-v1_3p","Hsa-Mir-154-P20-v2_3p","Hsa-Mir-154-P21_3p","Hsa-Mir-154-P22_3p","Hsa-Mir-154-P23_3p","Hsa-Mir-154-P24_3p","Hsa-Mir-154-P25_3p","Hsa-Mir-154-P26_3p","Hsa-Mir-154-P27_3p","Hsa-Mir-154-P28a_3p","Hsa-Mir-154-P28b_5p","Hsa-Mir-154-P28b_3p","Hsa-Mir-154-P29_3p","Hsa-Mir-154-P30_3p","Hsa-Mir-154-P36_3p","Hsa-Mir-155_5p","Hsa-Mir-181-P1a_5p","Hsa-Mir-181-P1b_5p","Hsa-Mir-181-P1c_5p","Hsa-Mir-181-P2a_5p","Hsa-Mir-181-P2b_5p","Hsa-Mir-181-P2c_5p","Hsa-Mir-184_3p","Hsa-Mir-185_5p","Hsa-Mir-186_5p","Hsa-Mir-187_3p","Hsa-Mir-188-P1_5p","Hsa-Mir-188-P2_5p","Hsa-Mir-188-P3_5p","Hsa-Mir-190-P1_5p","Hsa-Mir-190-P3_5p","Hsa-Mir-191_5p","Hsa-Mir-192-P1_5p","Hsa-Mir-192-P2_5p","Hsa-Mir-193-P1a_3p","Hsa-Mir-193-P1b_5p","Hsa-Mir-193-P2a_3p","Hsa-Mir-193-P2b_3p","Hsa-Mir-194-P1_5p","Hsa-Mir-194-P2_5p","Hsa-Mir-196-P1_5p","Hsa-Mir-196-P3_5p","Hsa-Mir-196-P4_5p","Hsa-Mir-197_3p","Hsa-Mir-199-P1-v1_3p","Hsa-Mir-199-P1-v2_3p","Hsa-Mir-199-P1-v3_3p","Hsa-Mir-199-P2-v1_3p","Hsa-Mir-199-P2-v2_3p","Hsa-Mir-199-P2-v3_3p","Hsa-Mir-199-P3-v1_3p","Hsa-Mir-199-P3-v2_3p","Hsa-Mir-199-P3-v3_3p","Hsa-Mir-202_5p","Hsa-Mir-203-v1_3p","Hsa-Mir-203-v2_3p","Hsa-Mir-204-P1_5p","Hsa-Mir-204-P2_5p","Hsa-Mir-205-P4_5p","Hsa-Mir-208-P1_3p","Hsa-Mir-208-P2_3p","Hsa-Mir-210_3p","Hsa-Mir-214-v1_3p","Hsa-Mir-214-v2_3p","Hsa-Mir-216-P2a_5p","Hsa-Mir-216-P2b_5p","Hsa-Mir-217-v1_5p","Hsa-Mir-217-v2_5p","Hsa-Mir-218-P2_5p","Hsa-Mir-218-P4_5p","Hsa-Mir-219-P2_5p","Hsa-Mir-219-P2_3p","Hsa-Mir-219-P2-as_5p","Hsa-Mir-219-P2-as_3p","Hsa-Mir-219-P3_3p","Hsa-Mir-221-P1a_3p","Hsa-Mir-221-P2a_5p","Hsa-Mir-221-P2a_3p","Hsa-Mir-223_3p","Hsa-Mir-224_5p","Hsa-Mir-296_5p","Hsa-Mir-320-P1_3p","Hsa-Mir-320-P2a_3p","Hsa-Mir-320-P2b_3p","Hsa-Mir-320-P3_3p","Hsa-Mir-320-P4_3p","Hsa-Mir-324_5p","Hsa-Mir-325_3p","Hsa-Mir-326_3p","Hsa-Mir-328_3p","Hsa-Mir-330-v1_3p","Hsa-Mir-330-v2_3p","Hsa-Mir-331_3p","Hsa-Mir-335_5p","Hsa-Mir-335_3p","Hsa-Mir-337-as_5p","Hsa-Mir-337-as_3p","Hsa-Mir-337-v1_5p","Hsa-Mir-337-v1_3p","Hsa-Mir-337-v2_5p","Hsa-Mir-337-v2_3p","Hsa-Mir-338-P2_3p","Hsa-Mir-338-P2-as_5p","Hsa-Mir-338-P2-as_3p","Hsa-Mir-339_5p","Hsa-Mir-339_3p","Hsa-Mir-340_5p","Hsa-Mir-342_3p","Hsa-Mir-345_5p","Hsa-Mir-346_5p","Hsa-Mir-361-v1_5p","Hsa-Mir-361-v2_3p","Hsa-Mir-362-P1_5p","Hsa-Mir-362-P2-v1_3p","Hsa-Mir-362-P2-v2_3p","Hsa-Mir-362-P3_3p","Hsa-Mir-362-P4_3p","Hsa-Mir-362-P5_5p","Hsa-Mir-370_3p","Hsa-Mir-374-P1_5p","Hsa-Mir-374-P1_3p","Hsa-Mir-374-P2_5p","Hsa-Mir-375_3p","Hsa-Mir-376-P1_3p","Hsa-Mir-376-P2_3p","Hsa-Mir-376-P3_3p","Hsa-Mir-376-P4_3p","Hsa-Mir-376-P5_3p","Hsa-Mir-378_3p","Hsa-Mir-383-v1_5p","Hsa-Mir-383-v2_5p","Hsa-Mir-423_5p","Hsa-Mir-423_3p","Hsa-Mir-425_5p","Hsa-Mir-430-P1_3p","Hsa-Mir-430-P2_3p","Hsa-Mir-430-P3_3p","Hsa-Mir-430-P4_3p","Hsa-Mir-430-P5_5p","Hsa-Mir-430-P5_3p","Hsa-Mir-430-P6_3p","Hsa-Mir-430-P7_3p","Hsa-Mir-430-P8a_3p","Hsa-Mir-430-P8b_3p","Hsa-Mir-430-P9a_5p","Hsa-Mir-430-P9a_3p","Hsa-Mir-430-P9b_5p","Hsa-Mir-430-P9b_3p","Hsa-Mir-430-P10_3p","Hsa-Mir-430-P11_3p","Hsa-Mir-430-P12_5p","Hsa-Mir-430-P13_5p","Hsa-Mir-430-P14_3p","Hsa-Mir-430-P15_5p","Hsa-Mir-430-P16_3p","Hsa-Mir-430-P17_5p","Hsa-Mir-430-P18_5p","Hsa-Mir-430-P18_3p","Hsa-Mir-430-P19_3p","Hsa-Mir-430-P21_3p","Hsa-Mir-430-P22_3p","Hsa-Mir-430-P23_3p","Hsa-Mir-430-P24_3p","Hsa-Mir-430-P25_3p","Hsa-Mir-430-P26_3p","Hsa-Mir-430-P27a_3p","Hsa-Mir-430-P27b_3p","Hsa-Mir-430-P29a_3p","Hsa-Mir-430-P29b_3p","Hsa-Mir-430-P31a_5p","Hsa-Mir-430-P31b_5p","Hsa-Mir-430-P33_5p","Hsa-Mir-430-P34_5p","Hsa-Mir-430-P35a_3p","Hsa-Mir-430-P35b_3p","Hsa-Mir-430-P35c_3p","Hsa-Mir-430-P38_3p","Hsa-Mir-430-P39_3p","Hsa-Mir-430-P41_3p","Hsa-Mir-430-P42_3p","Hsa-Mir-430-P43_5p","Hsa-Mir-430-P43_3p","Hsa-Mir-430-P44_5p","Hsa-Mir-430-P45_5p","Hsa-Mir-430-P46_5p","Hsa-Mir-430-P47_5p","Hsa-Mir-430-P48_5p","Hsa-Mir-430-P49_5p","Hsa-Mir-430-P50_5p","Hsa-Mir-431_5p","Hsa-Mir-431_3p","Hsa-Mir-432_5p","Hsa-Mir-433_3p","Hsa-Mir-448_5p","Hsa-Mir-448_3p","Hsa-Mir-450-P1_5p","Hsa-Mir-450-P2_5p","Hsa-Mir-450-P3_5p","Hsa-Mir-451_5p","Hsa-Mir-452-v1_5p","Hsa-Mir-452-v2_5p","Hsa-Mir-455_5p","Hsa-Mir-459_5p","Hsa-Mir-483_5p","Hsa-Mir-486_5p","Hsa-Mir-488_3p","Hsa-Mir-489-v1_3p","Hsa-Mir-489-v2_3p","Hsa-Mir-490_5p","Hsa-Mir-491_5p","Hsa-Mir-493_5p","Hsa-Mir-499_5p","Hsa-Mir-504-v1_5p","Hsa-Mir-504-v2_5p","Hsa-Mir-505-v1_5p","Hsa-Mir-505-v2_5p","Hsa-Mir-506-P1_5p","Hsa-Mir-506-P1_3p","Hsa-Mir-506-P2_3p","Hsa-Mir-506-P3_3p","Hsa-Mir-506-P4a1_3p","Hsa-Mir-506-P4a2_3p","Hsa-Mir-506-P4b_3p","Hsa-Mir-506-P5_5p","Hsa-Mir-506-P6a1_5p","Hsa-Mir-506-P6a2_5p","Hsa-Mir-506-P6b_5p","Hsa-Mir-506-P6c_5p","Hsa-Mir-506-P7a_3p","Hsa-Mir-506-P7b_3p","Hsa-Mir-506-P7c_3p","Hsa-Mir-506-P8_5p","Hsa-Mir-506-P9_5p","Hsa-Mir-506-P10_5p","Hsa-Mir-506-P11_3p","Hsa-Mir-506-P12_3p","Hsa-Mir-506-P13_5p","Hsa-Mir-506-P13_3p","Hsa-Mir-506-P29_3p","Hsa-Mir-511_5p","Hsa-Mir-511_3p","Hsa-Mir-541_5p","Hsa-Mir-541_3p","Hsa-Mir-542-v1_3p","Hsa-Mir-542-v2_3p","Hsa-Mir-544_3p","Hsa-Mir-550-P1_3p","Hsa-Mir-550-P2a_3p","Hsa-Mir-550-P2b_3p","Hsa-Mir-551-P1_3p","Hsa-Mir-551-P2_3p","Hsa-Mir-552_5p","Hsa-Mir-552_3p","Hsa-Mir-556_5p","Hsa-Mir-556_3p","Hsa-Mir-561_5p","Hsa-Mir-574_3p","Hsa-Mir-576_5p","Hsa-Mir-577_5p","Hsa-Mir-579_3p","Hsa-Mir-580_3p","Hsa-Mir-582-v1_5p","Hsa-Mir-582-v2_5p","Hsa-Mir-584_5p","Hsa-Mir-585_3p","Hsa-Mir-589_5p","Hsa-Mir-590_3p","Hsa-Mir-592_5p","Hsa-Mir-598_3p","Hsa-Mir-599_5p","Hsa-Mir-605-v1_3p","Hsa-Mir-605-v2_3p","Hsa-Mir-615_3p","Hsa-Mir-616_5p","Hsa-Mir-624_5p","Hsa-Mir-628_5p","Hsa-Mir-628_3p","Hsa-Mir-629_5p","Hsa-Mir-642_5p","Hsa-Mir-651_5p","Hsa-Mir-652_3p","Hsa-Mir-653_3p","Hsa-Mir-670_3p","Hsa-Mir-671_5p","Hsa-Mir-671_3p","Hsa-Mir-675_3p","Hsa-Mir-676_3p","Hsa-Mir-744_5p","Hsa-Mir-760_3p","Hsa-Mir-766_3p","Hsa-Mir-769_5p","Hsa-Mir-770_5p","Hsa-Mir-873-v1_5p","Hsa-Mir-873-v1_3p","Hsa-Mir-873-v2_5p","Hsa-Mir-874_3p","Hsa-Mir-875_5p","Hsa-Mir-876_3p","Hsa-Mir-877_5p","Hsa-Mir-885_5p","Hsa-Mir-887_5p","Hsa-Mir-887_3p","Hsa-Mir-935-v1_3p","Hsa-Mir-935-v2_3p","Hsa-Mir-944_3p","Hsa-Mir-1180_3p","Hsa-Mir-1247_5p","Hsa-Mir-1249_3p","Hsa-Mir-1251_5p","Hsa-Mir-1252_5p","Hsa-Mir-1266_5p","Hsa-Mir-1269_3p","Hsa-Mir-1270_5p","Hsa-Mir-1271_5p","Hsa-Mir-1271_3p","Hsa-Mir-1276_5p","Hsa-Mir-1277_5p","Hsa-Mir-1278_3p","Hsa-Mir-1286_3p","Hsa-Mir-1287_5p","Hsa-Mir-1294_5p","Hsa-Mir-1295_3p","Hsa-Mir-1296_5p","Hsa-Mir-1298_5p","Hsa-Mir-1301_3p","Hsa-Mir-1306_5p","Hsa-Mir-1307_5p","Hsa-Mir-1323_5p","Hsa-Mir-1343_3p","Hsa-Mir-1908_5p","Hsa-Mir-1910_3p","Hsa-Mir-1911_5p","Hsa-Mir-1912_3p","Hsa-Mir-1913_5p","Hsa-Mir-1913_3p","Hsa-Mir-1914_5p","Hsa-Mir-2114_5p","Hsa-Mir-2114_3p","Hsa-Mir-2115_3p","Hsa-Mir-2116_5p","Hsa-Mir-2116_3p","Hsa-Mir-2276_5p","Hsa-Mir-2276_3p","Hsa-Mir-2355_5p","Hsa-Mir-2355_3p","Hsa-Mir-2467_5p","Hsa-Mir-3059_5p","Hsa-Mir-3085_3p","Hsa-Mir-3126_5p","Hsa-Mir-3127_5p","Hsa-Mir-3136_5p","Hsa-Mir-3138_3p","Hsa-Mir-3140_3p","Hsa-Mir-3144-v1_5p","Hsa-Mir-3144-v2_5p","Hsa-Mir-3145_3p","Hsa-Mir-3146_3p","Hsa-Mir-3173_5p","Hsa-Mir-3174_5p","Hsa-Mir-3180-P1_3p","Hsa-Mir-3180-P2_3p","Hsa-Mir-3180-P3_3p","Hsa-Mir-3187_3p","Hsa-Mir-3194_5p","Hsa-Mir-3200_3p","Hsa-Mir-3611_3p","Hsa-Mir-3613_5p","Hsa-Mir-3617_5p","Hsa-Mir-3619_5p","Hsa-Mir-3622_5p","Hsa-Mir-3622_3p","Hsa-Mir-3663_3p","Hsa-Mir-3690_5p","Hsa-Mir-3909_3p","Hsa-Mir-3924_3p","Hsa-Mir-3934_5p","Hsa-Mir-3934_3p","Hsa-Mir-3942_5p","Hsa-Mir-3942_3p","Hsa-Mir-3944_3p","Hsa-Mir-4424_3p","Hsa-Mir-4524_3p","Hsa-Mir-4637_3p","Hsa-Mir-4661_5p","Hsa-Mir-4662_5p","Hsa-Mir-4664_5p","Hsa-Mir-4664_3p","Hsa-Mir-4677_3p","Hsa-Mir-4731_5p","Hsa-Mir-4731_3p","Hsa-Mir-4766_5p","Hsa-Mir-4766_3p","Hsa-Mir-5579_5p","Hsa-Mir-5581_5p","Hsa-Mir-5581_3p","Hsa-Mir-5586_5p","Hsa-Mir-5683_5p","Hsa-Mir-5687_5p","Hsa-Mir-5687_3p","Hsa-Mir-6715_3p","Hsa-Mir-7151_5p","Hsa-Mir-7151_3p","Hsa-Mir-7854_3p","Hsa-Novel-1-v1_3p","Hsa-Novel-2_5p","Hsa-Novel-2_3p","Hsa-Novel-3_5p"
"ERR409886",18482,47783,208285,208285,208285,129115,129115,27498,21535,7913,16405,45477,89,1,89,1932,1932,1932,381,46,389,349,466,162369,8029,162369,8029,162369,8029,33274,2402,3269,551,84391,22426,52166,181833,85271,85271,3166,363,352,2111,36647,37811,20,5760,662,164,473,254,104,28,884,163,3226,399,1216,1216,21918,4210,9845,3637,3329,3329,1006516,30890,1006516,163575,11134,1438,14574,29328,6468,1337,6548,2989,6548,2989,43894,11586,79701,187929,81149,14185,36070,36070,949,293,433,323,967,79,17,589,7,8,7,0,16,22290,22290,78218,1,2416,5989,994,2462,10,8,4,25,217,7399,1483,590,40121,31860,40121,31860,7270,35489,35489,78,78,5,41977,52733,41977,52733,41977,52733,24196,106512,25090,128705,72327,72327,2106,3709,546,4454,518,3843,1086,190,287,6120,302,753,1559,508,52,9,1559,508,720,1351,1351,8,9448,1723,1608,2194,254,11997,11997,6497,4501,8186,1835,76,367,2979,620467,1162,1282,7301,17722,967,4,5,11049,191,3009,10014,1904,11572,11572,268,20,85,59,1103,2084,1134,502,502,367,493,450,432,27,209,11651,412,7405,9289,20935,92,840,462,2164,87,424,19,18,1622,309,4200,62,379,515,2947,18,355,4,0,149,3582,695,392597,392597,43191,22462,22462,5225,141,285,25940,79,26,1276,1129,2275,18,119626,902,8147,410,177,74,465,574,580,16,12,19,6654,3161,3637,359,3161,3637,359,3161,3637,359,0,370,90,174,18936,9,56,1,456,79,121,0,0,0,3,3644,3644,2830,1575,354,2139,4,11573,888,17482,869,122,20,2912,284,284,74,79,597,0,73,327,750,960,920,4664,1975,2,10,10,35,24,63,21870,21979,248,96,1043,8654,6748,1105,247,2537,1202,196,1259,987,711,213,23,609,804,1278,863,373,217,37,217,911,4,7172,266,313,3003,8642,2484,2,1,7,6,0,3,1,4,0,0,3,0,3,0,4,3,0,2,0,8,23,2,2,1,3,0,2,2,1,0,0,0,0,0,0,127,127,20,1,2,2,3,0,0,30,0,0,6,0,0,0,0,3,8,8,16,27,1650,1122,3,7,98,98,181,9006,41,36,263,0,84,21802,607,25,3,26,70,36,132,672,582,5,0,0,1,0,5,13,13,13,0,1,1,0,2,2,2,2,0,6,1,7,0,0,1,0,8,6,9,12,42,36,6,57,57,57,6,63,2,2,2,0,90,1141,70,1294,0,6,280,125,610,3,333,83,188,2777,0,2,0,10,17,6,847,372,63,3,33,178,1,62,78,443,7,1,4015,21,274,20009,470,1630,309,1077,2891,2,38,192,1026,22,841,944,975,2,1489,126,914,77,19,0,3,7,799,0,9,76,1,36,17,1,1,2648,284,370,49,9072,1,32,21,0,40,18,1,4,1,9,6,9,1,5,0,1,38,1,163,74,8,0,13,1,2,0,0,0,0,0,4,1,16,16,16,0,6,719,14,143,2,1,8,16,0,4,74,0,1,0,1,0,0,10,0,0,0,44,0,1,33,0,1,0,2,0,0,4,23,427,0,0,1,0,3,4,0,1,2,1
"ERR409894",25295,74145,314056,314056,314056,196868,196868,44708,38319,12313,25733,73578,80,0,80,2544,2544,2544,60,12,52,47,23,296685,5669,296685,5669,296685,5669,61620,4875,1558,331,41100,14486,30986,81252,66568,66568,4681,473,316,3132,43675,45066,17,4206,806,114,446,364,131,17,1169,170,4443,296,1181,1181,16810,3742,11247,2053,3471,3471,1077711,36069,1077711,251199,4538,962,10434,38256,15234,1483,9233,3091,9233,3091,40611,13395,112919,181079,77627,20833,38564,38564,1215,336,1173,523,1588,102,15,992,11,20,2,0,18,35052,35052,64729,0,1835,7599,1128,4416,11,9,2,24,9,1411,50,26,49727,59678,49727,59678,12143,60887,60887,146,146,24,59511,69508,59511,69508,59511,69508,13765,73971,14378,181488,87393,87393,2443,7258,857,3754,448,6669,1345,93,269,10423,319,828,1134,372,39,2,1134,372,1886,1296,1296,8,16570,2736,1479,2334,206,28543,28543,8033,4699,8944,1127,44,190,1110,197946,893,1024,3502,25612,1136,15,16,6976,173,3722,16591,1010,12486,12486,349,73,118,109,2897,4897,1953,747,747,887,868,455,1055,34,233,7366,1364,27279,17203,36872,35,1618,1180,3475,225,637,42,75,2579,264,2381,142,595,1042,5190,79,896,8,0,303,10352,306,1210291,1210291,62245,81344,81344,7834,212,430,25963,115,26,1259,1342,2729,17,135192,941,8565,694,243,67,359,519,532,12,11,16,4628,1120,1313,122,1120,1313,122,1120,1313,122,0,669,143,13,12255,5,126,0,575,15,19,0,9,4,3,5260,5260,10450,8358,2101,7874,2,16851,496,23335,301,31,33,3306,433,433,113,116,1079,0,93,471,1467,1740,1499,3550,1611,2,3,3,47,16,69,60609,60760,352,154,1462,32764,6652,1518,385,2464,1797,137,985,829,292,114,22,979,940,2096,931,1010,278,35,278,1288,0,5042,699,901,2677,6997,4201,7,4,8,6,0,0,0,4,1,1,6,0,6,0,3,0,0,1,0,18,20,1,1,2,2,2,0,0,0,0,0,0,0,1,1,123,123,2,1,2,2,4,2,2,16,0,0,14,0,0,0,1,0,9,9,76,90,3853,2099,0,6,99,99,210,17440,7,5,161,1,11,20637,1413,30,7,91,175,59,227,607,568,10,2,0,1,0,10,13,13,13,0,4,4,1,7,16,16,16,0,0,0,5,0,0,1,0,7,2,11,66,112,51,8,29,29,29,2,76,5,0,1,1,25,813,60,3218,2,3,469,238,2573,1,597,92,615,4342,0,0,0,8,18,1,1402,444,54,7,57,253,6,136,151,705,1,7,8443,48,226,59904,413,3923,880,2508,4247,0,59,464,1511,24,1237,1708,1775,0,2620,103,1958,183,15,4,6,10,225,2,13,75,0,55,55,1,0,2771,95,435,80,13812,5,59,56,0,9,3,1,4,3,4,2,16,0,7,1,9,38,3,352,163,37,0,12,0,0,0,0,0,0,2,6,0,8,8,8,5,0,1544,11,384,2,0,18,10,5,0,94,0,0,0,2,0,2,11,2,0,2,69,0,0,104,2,0,1,0,0,0,7,6,497,0,1,0,9,20,4,2,2,0,1
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"ERR409888",20686,53365,320237,320237,320237,154352,154352,56350,35260,10690,24807,53362,33,0,33,1368,1368,1368,66,6,44,21,16,160209,4621,160209,4621,160209,4621,603,38,773,135,55847,11674,53908,87696,61217,61217,4751,527,386,1869,56121,57923,18,4529,934,62,510,403,222,15,1176,93,6879,358,1284,1284,14586,3041,28252,2087,3465,3465,839104,33080,839104,422947,4270,894,8432,57885,20470,1302,3688,2158,3688,2158,26618,8055,77062,101046,55954,13633,26101,26101,873,354,1031,166,683,177,56,1336,4,6,2,0,10,41615,41615,33066,2,1010,8566,735,6452,12,14,3,17,13,811,41,14,31455,39013,31455,39013,9413,50436,50436,75,75,2,17624,22097,17624,22097,17624,22097,13641,38517,14052,86092,43830,43830,2068,4274,1014,2841,467,6222,1645,124,371,11746,205,779,785,231,40,2,785,231,802,805,805,16,6748,971,546,1363,174,14740,14740,6313,5682,10425,720,38,145,1183,78666,259,309,2082,25018,1047,11,15,2906,120,3632,9871,871,4654,4654,214,23,63,57,1385,2505,1068,515,515,362,405,212,404,11,147,7493,524,12997,7498,18143,25,720,538,1735,138,568,13,35,1200,260,1705,60,313,429,2645,25,361,3,0,136,4378,340,1838301,1838301,41764,152459,152459,5508,224,290,27699,46,24,983,1068,1030,9,100751,752,7911,1072,139,147,533,397,412,4,0,0,6048,433,523,58,433,523,58,433,523,58,0,469,116,506,10508,7,37,0,455,5,10,3,6,1,1,2391,2391,13625,23548,5949,8819,4,9217,516,13915,403,7,35,4107,937,937,303,309,855,0,66,287,1817,2179,1294,2752,1210,1,7,7,26,12,36,108683,109010,824,147,740,25939,6245,969,315,2225,1719,109,819,695,282,89,9,517,750,1190,1147,499,161,18,161,576,4,4638,586,734,2498,6309,2966,0,0,0,3,6,3,14,18,0,0,2,0,2,0,0,1,0,0,0,3,6,0,0,1,0,0,0,0,0,1,0,1,1,0,0,24,24,4,1,0,0,0,0,0,15,0,0,2,0,0,0,0,0,1,1,29,25,2337,1155,1,5,53,53,112,10767,9,5,126,0,6,7143,501,23,1,39,99,43,211,357,314,10,3,0,0,0,11,14,14,14,0,1,1,0,5,1,1,1,0,0,0,4,0,0,0,0,4,1,14,15,60,31,6,38,38,38,9,54,4,5,9,2,24,2179,87,11631,1,6,188,106,7185,2,441,75,319,3627,0,0,0,0,33,2,745,311,18,58,19,178,3,120,57,326,4,7,4876,40,190,39144,243,1567,331,985,3021,0,39,317,1216,26,1056,935,994,0,1199,28,1052,76,18,3,6,32,603,2,14,54,2,29,72,3,1,2724,119,296,51,3220,2,32,21,0,5,6,0,2,1,8,4,10,2,19,2,16,73,2,101,59,11,0,13,3,3,1,0,0,0,0,16,4,9,9,9,0,0,1284,3,81,10,0,5,7,14,14,85,0,3,0,0,0,0,5,2,1,1,30,0,0,98,0,0,0,0,0,0,9,6,213,0,0,1,7,10,2,0,0,3,0
"ERR409883",50124,117559,287216,287216,287216,150966,150966,100340,51970,9076,78984,50759,302,0,302,40367,40367,40367,181,26,225,201,59,1119735,27361,1119735,27361,1119735,27361,5018,316,3471,687,468097,64336,91004,268350,354990,354990,4998,284,895,4963,42614,44550,96,13652,1311,292,2499,2068,649,105,1269,469,10365,817,5402,5402,33324,10176,13571,5281,23971,23971,1911607,44636,1911607,407620,23050,1703,19193,33314,46720,8424,35669,11189,35669,11189,171798,57777,349596,563598,335059,32911,69221,69221,2883,491,1586,3812,4965,74,21,802,11,14,11,1,63,39084,39084,76677,0,3661,12219,3110,4234,19,17,16,268,56,3398,2523,1335,147985,189832,147985,189832,37333,208477,208477,571,571,35,498334,612748,498334,612748,498334,612748,62043,168531,63733,2090432,269080,269080,20258,64162,4986,13149,1937,7467,934,340,1205,20602,3495,2896,7158,2419,991,58,7158,2419,11113,3078,3078,65,47726,7408,14128,29901,1870,74909,74909,40377,19877,28272,3074,131,581,3413,1060230,1685,1921,10765,156907,14390,67,77,16279,1124,32150,77904,1688,51619,51619,1586,379,256,276,9602,13797,7483,4254,4254,704,1731,2030,9343,318,493,26271,6867,40611,18954,35032,817,4894,1261,6952,343,4678,122,152,1793,1535,7937,1151,780,5730,5347,169,1856,31,3,1592,29631,951,427957,427957,100468,66958,66958,20062,957,2401,52335,968,69,6524,3906,4684,25,650151,1497,23833,3063,529,471,1168,856,875,5,0,0,15747,2042,2337,221,2042,2337,221,2042,2337,221,1,3725,689,1400,50953,17,319,0,1971,87,137,9,57,10,11,20344,20344,22926,10814,5018,16382,3,61170,2346,87707,538,7,24,23708,4792,4792,1507,1515,4305,0,497,3006,4138,6203,10836,19210,6120,8,36,36,71,80,133,57358,57586,701,1067,3959,60034,63126,2948,1604,7940,5494,1473,2578,1889,1300,582,84,11665,2101,2616,1650,1248,2230,210,2230,11983,18,28375,1408,1585,6434,11302,11004,2,0,5,9,10,1,5,35,2,2,6,0,6,0,6,7,3,11,3,30,109,11,11,1,2,1,6,6,0,0,2,0,0,4,4,532,532,21,4,3,3,0,10,10,63,0,3,50,2,3,3,0,12,12,12,140,531,14287,7284,24,89,280,280,351,16557,24,27,941,0,72,48819,5988,36,1,661,2194,145,641,5311,4509,2,0,0,3,2,45,108,108,108,4,3,3,0,2,13,13,13,1,7,6,5,0,0,0,0,8,16,99,97,507,326,42,101,101,101,13,438,22,35,7,4,246,1742,48,3985,3,13,1169,1035,3764,14,1101,78,1212,8978,0,0,0,1,41,5,1220,750,467,14,131,1110,21,266,1341,1691,22,7,31197,422,1714,379207,2475,32833,10260,21435,34460,0,320,840,4514,122,3861,3534,3761,3,5786,255,2912,2242,63,18,42,39,174,12,69,147,0,512,114,2,14,16093,223,2379,219,96360,2,192,70,1,29,57,7,55,8,38,19,36,4,12,0,7,91,19,1664,386,142,3,46,0,0,0,0,0,0,0,4,2,158,158,158,6,10,5946,72,632,18,4,62,18,6,26,159,0,28,2,4,0,12,24,1,0,0,39,3,0,250,1,4,0,2,1,2,26,21,5035,3,3,0,3,5,8,0,0,1,3
"ERR409900",114606,271460,1258967,1258967,1258967,802974,802974,174495,142624,55680,97566,310818,449,2,449,20439,20439,20439,164,45,140,90,82,1160019,26481,1160019,26481,1160019,26481,1784,136,2827,530,223093,54743,131887,378490,235292,235292,16596,1905,1287,13171,177881,182861,44,23195,2793,575,1619,997,455,66,4785,876,19463,772,3276,3276,49412,16041,39170,8280,13975,13975,2066195,124098,2066195,999775,18861,3739,36351,153639,45029,5772,28289,10093,28289,10093,127958,41609,403791,585382,311134,71196,161776,161776,6805,1286,3451,1312,7548,408,84,2460,20,34,14,0,36,108578,108578,300449,0,7367,27213,4329,18039,51,37,36,118,58,3342,186,103,134743,168965,134743,168965,47477,252692,252692,535,535,69,150302,181910,150302,181910,150302,181910,46629,260473,49065,602166,333051,333051,12122,33485,4616,14620,2601,24631,5466,458,1361,60706,1678,5030,5166,1492,167,5,5166,1492,4476,9489,9489,148,58020,8347,5435,8629,1034,115714,115714,35419,19893,35601,4710,210,830,4827,497102,2252,2510,9263,134204,5168,23,29,19534,483,13217,53480,4833,43066,43066,1315,184,585,553,8569,14720,8291,2470,2470,3515,3396,1706,3964,70,1106,42961,4609,109192,64366,150646,123,5312,5111,14033,661,3690,215,296,10706,1698,10548,437,2398,3018,20627,214,3808,26,2,1150,33478,1115,4350668,4350668,228499,320102,320102,27194,689,2334,87752,665,101,3461,3804,10323,25,500036,3042,26345,2294,909,428,1924,1708,1737,9,1,3,20189,3434,4117,338,3434,4117,338,3434,4117,338,0,2537,615,1928,41926,32,318,1,1816,84,118,9,12,4,19,17955,17955,29759,32651,8088,21118,7,84282,3645,90702,1322,46,84,13284,1725,1725,672,681,3642,0,298,1480,5476,6658,5486,15309,8632,8,44,44,169,133,259,213137,213856,1507,336,4280,94473,28328,5058,1296,8817,5286,662,4059,3497,1270,386,91,4071,3045,5972,3084,2991,946,156,946,4217,18,21283,3045,3766,11448,27542,15958,11,1,4,10,5,1,8,17,0,0,10,0,10,0,9,4,1,2,0,21,55,2,2,3,4,0,0,0,1,0,4,1,1,2,2,192,192,9,1,6,6,0,2,2,50,0,3,12,2,1,1,2,3,9,9,367,316,12500,8927,23,37,395,395,763,62895,38,23,631,0,21,76857,4194,166,17,228,449,240,651,2017,1856,29,7,1,1,0,43,43,43,43,1,8,8,0,15,32,32,32,0,15,1,20,4,1,1,0,30,9,62,171,338,200,50,201,201,201,4,178,15,10,17,11,165,4232,262,17181,0,28,1533,714,11392,33,1782,303,2428,13519,1,0,0,2,81,15,4184,1386,164,50,146,694,10,887,646,2322,8,25,32413,158,1081,193939,1838,9853,2269,6412,14719,0,278,1590,7838,124,5040,6012,6372,5,9394,206,4752,975,97,8,24,53,1716,3,60,336,9,254,126,15,12,8235,482,2372,200,42483,1,315,261,1,6,39,2,4,5,19,19,52,4,29,2,14,282,12,951,664,73,3,49,5,0,1,1,0,2,2,26,10,52,52,52,4,7,5746,39,893,7,2,97,25,17,13,415,0,2,0,4,1,3,71,2,0,1,207,1,0,353,0,2,1,1,0,1,39,115,2549,0,0,1,22,16,16,0,1,2,1

Note: the “mature_counts.csv” needs to be transposed prior running the statistical analysis. This can be done either user the R script or using a script called “transpose_csv.py”.

Let’s copy the transpose_csv.py script to the working folder:

Code Block
cp /work/training/2024/smallRNAseq/scripts/transpose_csv.py .

The check how to use the script do the following:

Code Block
python transpose_csv.py --help
Code Block
usage: transpose_csv.py [-h] --input INPUT --output OUTPUT
Transpose a CSV file and generate a tab-delimited TXT file.
optional arguments:
  -h, --help       show this help message and exit
  --input INPUT    Input CSV file containing mature miRNA counts.
  --output OUTPUT  Output tab-delimited TXT file.

To transpose the initial “mature_counst.csv” file do the following:

Code Block
python transpose_csv.py --input mature_counts.csv --out mature_counts.txt

Let’s now print the transposed mature counts table:

Code Block
cat mature_counts.txt
Code Block
microRNA	ERR409886	ERR409894	ERR409882	ERR409895	ERR409889	ERR409899	ERR409879	ERR409885	ERR409881ERR409884	ERR409887	ERR409896	ERR409891	ERR409880	ERR409878	ERR409898	ERR409897	ERR409893	ERR409890ERR409892	ERR409888	ERR409883	ERR409900
Hsa-Let-7-P1b_5p	18482	25295	29486	51987	44172	26036	23604	46550	68044	13902	15724	33998	29525	24585	29935	15501	19495	39737	31807	20070	20686	50124	114606
Hsa-Let-7-P1c_5p	47783	74145	68342	118227	100021	66085	58426	128501	162724	39989	41506	84952	75858	65606	69995	41283	46267	86159	73537	62698	53365	117559	271460
Hsa-Let-7-P1d_5p	208285	314056	364624	531423	568283	289927	305660	546059	712891	182180	178203	421398	351589	318124	432959	186915	242780	417695	332584	249356	320237	287216	1258967
Hsa-Let-7-P2a1_5p	208285	314056	364624	531423	568283	289927	305660	546059	712891	182180	178203	421398	351589	318124	432959	186915	242780	417695	332584	249356	320237	287216	1258967
Hsa-Let-7-P2a2_5p	208285	314056	364624	531423	568283	289927	305660	546059	712891	182180	178203	421398	351589	318124	432959	186915	242780	417695	332584	249356	320237	287216	1258967
Hsa-Let-7-P2a3_5p	129115	196868	211190	425095	276482	223463	198784	376491	456871	127391	123392	276681	231752	191076	250199	126873	155636	288225	207846	133797	154352	150966	802974
Hsa-Let-7-P2b1_5p	129115	196868	211190	425095	276482	223463	198784	376491	456871	127391	123392	276681	231752	191076	250199	126873	155636	288225	207846	133797	154352	150966	802974
Hsa-Let-7-P2b2_5p	27498	44708	59417	75630	92339	43602	52115	59795	83857	27892	25678	45870	37230	46347	61357	25897	28396	62467	40131	28694	56350	100340	174495
Hsa-Let-7-P2b3_5p	21535	38319	50824	54780	45533	31044	30986	58479	80694	21549	21471	45473	42277	33601	48053	22597	25577	46855	39626	25804	35260	51970	142624
Hsa-Let-7-P2c1_5p	7913	12313	14861	27318	20819	14122	12397	22746	30556	7751	7833	17317	14405	11095	15326	8698	10109	19831	13528	9305	10690	9076	55680
Hsa-Let-7-P2c2_5p	16405	25733	35925	46007	40731	26046	31814	52245	48422	22131	16863	28264	28404	30137	36839	16318	21184	38978	27316	15145	24807	78984	97566
Hsa-Let-7-P2c3_5p	45477	73578	72845	123354	109175	74340	71170	128485	173832	46596	39681	98399	83319	68711	96580	47999	55668	104358	82526	41838	53362	50759	310818
Hsa-Mir-1-P1_3p	89	80	74	94	90	52	118	226	236	66	47	190	181	97	118	57	79	187	123	24	33	302	449
Hsa-Mir-1-P2_3p	1	0	0	0	2	0	0	0	4	2	0	0	1	0	0	0	0	22
Hsa-Mir-1-P3_3p	89	80	74	94	90	52	118	226	236	66	47	190	181	97	118	57	79	187	123	24	33	302	449
Hsa-Mir-7-P1_5p	1932	2544	2622	2601	3681	1065	2172	3395	7532	1447	851	3483	3044	2989	3100	1450	1927	4548	4122	1976	1368	40367	20439
Hsa-Mir-7-P2_5p	1932	2544	2622	2601	3681	1065	2172	3395	7532	1447	851	3483	3044	2989	3100	1450	1927	4548	4122	1976	1368	40367	20439
Hsa-Mir-7-P4_5p	1932	2544	2622	2601	3681	1065	2172	3395	7532	1447	851	3483	3044	2989	3100	1450	1927	4548	4122	1976	1368	40367	20439
Hsa-Mir-8-P1a_3p	381	60	131	86	128	64	50	98	111	35	49	64	51	47	121	46	462	86	139	66	181	164
Hsa-Mir-8-P1b_3p	46	12	9	29	27	21	11	26	38	7	5	26	21	25	18	16	115	16	25	6	26	45
Hsa-Mir-8-P2a_3p	389	52	77	46	182	48	77	67	68	36	39	23	39	36	117	43	347	51	44	44	225	140
Hsa-Mir-8-P2b_3p	349	47	40	64	73	33	45	93	83	34	25	40	48	50	56	20	435	49	81	21	201	90
Hsa-Mir-8-P3a_3p	466	23	61	25	63	23	32	38	29	16	19	25	19	44	76	20	126	40	27	16	59	82
Hsa-Mir-9-P1_5p	162369	296685	325973	351690	348083	202181	244049	538146	505079	196178	145191	338071	320620	290747	375141	143367	221771	352483	313212	190994	160209	1119735	1160019
Hsa-Mir-9-P1_3p	8029	5669	8439	8239	12697	5564	6070	13606	11707	4898	4103	7300	7883	8820	7685	4557	5025	7405	9454	10681	4621	27361	26481
Hsa-Mir-9-P2_5p	162369	296685	325973	351690	348083	202181	244049	538146	505079	196178	145191	338071	320620	290747	375141	143367	221771	352483	313212	190994	160209	1119735	1160019
Hsa-Mir-9-P2_3p	8029	5669	8439	8239	12697	5564	6070	13606	11707	4898	4103	7300	7883	8820	7685	4557	5025	7405	9454	10681	4621	27361	26481
Hsa-Mir-9-P3_5p	162369	296685	325973	351690	348083	202181	244049	538146	505079	196178	145191	338071	320620	290747	375141	143367	221771	352483	313212	190994	160209	1119735	1160019
Hsa-Mir-9-P3_3p	8029	5669	8439	8239	12697	5564	6070	13606	11707	4898	4103	7300	7883	8820	7685	4557	5025	7405	9454	10681	4621	27361	26481
Hsa-Mir-10-P1b-v1_5p	33274	61620	590	20151	10343	7551	745	6828	33515	6965	653	1120	1043	46880	1104	315	30186	937	12711	12178	603	5018	1784
Hsa-Mir-10-P1b-v2_5p	2402	4875	39	1621	815	621	53	446	2722	504	47	90	75	3480	84	26	2067	70	916	810	38	316	136
Hsa-Mir-10-P1c-v1_5p	3269	1558	257	1767	896	405	556	6232	33628	726	1175	1313	599	2808	1498	287	686	1232	1025	3019	773	3471	2827
Hsa-Mir-10-P1c-v2_5p	551	331	40	374	173	69	81	906	6418	106	194	246	98	551	288	45	125	230	192	549	135	687	530
Hsa-Mir-10-P2b_5p	84391	41100	76456	45448	114204	53057	39073	64175	83922	43204	32872	38187	55062	69417	39121	25110	22826	65593	70976	58992	55847	468097	223093
Hsa-Mir-10-P2c_5p	22426	14486	15965	16006	25485	14763	13513	23895	33096	12728	9571	12327	15968	20179	13461	8407	8378	15562	18562	24577	11674	64336	54743
Hsa-Mir-10-P2d_5p	52166	30986	79481	28319	107620	40535	37070	60082	63987	30825	18025	28735	39781	44661	44899	20461	14628	41072	40955	38033	53908	91004	131887
Hsa-Mir-10-P3b_5p	181833	81252	138889	126317	224474	111938	83880	192590	206640	68002	66334	106078	137101	131928	88529	64913	62802	124192	140567	180845	87696	268350	378490
Hsa-Mir-10-P3c_5p	85271	66568	79205	77580	124928	63693	61766	105925	140562	44846	40183	57630	69745	78369	66116	41947	36981	73506	80308	103672	61217	354990	235292
Hsa-Mir-10-P3d_5p	85271	66568	79205	77580	124928	63693	61766	105925	140562	44846	40183	57630	69745	78369	66116	41947	36981	73506	80308	103672	61217	354990	235292
Hsa-Mir-15-P1a_5p	3166	4681	7002	12518	12407	5151	5486	7931	9893	3651	3288	5614	4055	5599	9213	3567	3704	5804	5146	4126	4751	4998	16596
Hsa-Mir-15-P1b_5p	363	473	516	719	857	312	563	822	1007	471	295	474	465	715	823	300	306	478	376	418	527	284	1905
Hsa-Mir-15-P1c_5p	352	316	560	205	511	120	250	567	557	282	115	277	305	289	572	111	197	282	297	354	386	895	1287
Hsa-Mir-15-P1d_5p	2111	3132	2437	6701	3326	3637	2450	3479	7965	2261	2204	3698	2989	3168	2864	1994	2213	4398	3161	3333	1869	4963	13171
Hsa-Mir-15-P2a_5p	36647	43675	49659	106970	83889	50438	48372	73527	111687	40310	39428	55347	45421	67394	73290	34801	35131	54599	42388	49614	56121	42614	177881
Hsa-Mir-15-P2b_5p	37811	45066	51704	108410	86937	51359	49837	76507	114249	41613	40251	57207	47166	69275	76485	35712	36475	55968	43780	51047	57923	44550	182861
Hsa-Mir-15-P2c_5p	20	17	8	17	22	5	10	22	34	13	3	9	17	17	17	10	911	20	16	18	96	44
Hsa-Mir-15-P2d_5p	5760	4206	4855	8138	6641	3753	4989	8525	12105	6397	3595	6472	6607	8438	7082	3257	4859	5490	5432	6714	4529	13652	23195
Hsa-Mir-17-P1a_5p	662	806	1091	822	1971	548	945	1079	1495	629	445	681	666	1242	1301	537	561	805	659	590	934	1311	2793
Hsa-Mir-17-P1c_5p	164	114	99	159	191	83	71	125	304	88	43	104	83	249	128	63	128	131	146	82	62	292	575
Hsa-Mir-17-P1d_5p	473	446	857	1033	1318	436	573	866	850	412	400	554	355	731	853	413	358	549	559	439	510	2499	1619
Hsa-Mir-17-P1d_3p	254	364	419	814	579	381	324	428	490	226	277	302	229	350	391	218	262	349	299	189	403	2068	997
Hsa-Mir-17-P2a_5p	104	131	326	101	403	76	194	137	174	97	65	98	104	168	327	85	7115	102	69	222	649	455
Hsa-Mir-17-P2c_5p	28	17	29	18	48	18	13	20	34	13	11	20	18	41	27	7	112	30	16	15	105	66
Hsa-Mir-17-P4a_5p	884	1169	1615	1036	3172	855	1317	1632	2220	902	461	1097	1199	1836	2163	673	869	1156	1086	649	1176	1269	4785
Hsa-Mir-17-P4c_5p	163	170	144	215	257	111	84	168	469	124	62	173	136	309	189	67	158	214	209	88	93	469	876
Hsa-Mir-17-P4d_5p	3226	4443	6901	8262	9831	3914	5416	5275	7821	3750	2768	4263	3474	5608	7290	2739	2781	5811	3821	2346	6879	10365	19463
Hsa-Mir-19-P1a_3p	399	296	468	307	1082	232	368	473	318	305	150	237	298	505	542	253	259	247	219	346	358	817	772
Hsa-Mir-19-P2a_3p	1216	1181	1482	1046	3082	766	1097	1656	1466	1147	539	878	1054	1708	1773	788	912	884	964	1312	1284	5402	3276
Hsa-Mir-19-P2c_3p	1216	1181	1482	1046	3082	766	1097	1656	1466	1147	539	878	1054	1708	1773	788	912	884	964	1312	1284	5402	3276
Hsa-Mir-21_5p	21918	16810	24378	23678	33803	20854	17441	28742	25992	11326	7940	22008	20342	16238	29409	10725	13724	17677	18528	13654	14586	33324	49412
Hsa-Mir-22-P1b_3p	4210	3742	6526	9282	6428	4826	3693	7321	10360	2276	2958	5011	4172	3652	4830	3335	2297	6177	6017	5602	3041	10176	16041
Hsa-Mir-23-P2_3p	9845	11247	17222	11499	14994	6013	11852	14345	16971	10756	9108	12375	14864	12411	28226	9190	7116	12957	8092	18031	28252	13571	39170
Hsa-Mir-23-P3_3p	3637	2053	2551	3093	2989	1923	1912	4189	4722	1657	1310	2482	2937	3303	3081	1396	1721	2606	2176	3275	2087	5281	8280
Hsa-Mir-24-P2_3p	3329	3471	5206	2510	6773	2039	4549	4608	6172	2715	1544	2830	2943	4085	5937	3148	1886	3436	3320	3630	3465	23971	13975
Hsa-Mir-24-P3_3p	3329	3471	5206	2510	6773	2039	4549	4608	6172	2715	1544	2830	2943	4085	5937	3148	1886	3436	3320	3630	3465	23971	13975
Hsa-Mir-26-P1_5p	1006516	1077711	912248	890275	1126662	568618	648819	1234738	1306207	444239	468744	808800	928138	992232	1475541	523073	559600	692502	880724	1170207	839104	1911607	2066195
Hsa-Mir-26-P2_5p	30890	36069	41809	42091	67728	28058	37424	71349	64732	24388	18438	40245	36161	47704	56249	22951	26013	36812	34280	33901	33080	44636	124098
Hsa-Mir-26-P4_5p	1006516	1077711	912248	890275	1126662	568618	648819	1234738	1306207	444239	468744	808800	928138	992232	1475541	523073	559600	692502	880724	1170207	839104	1911607	2066195
Hsa-Mir-27-P2_3p	163575	251199	483786	330699	646255	237548	386589	340718	456521	233163	136266	283426	222559	251724	638569	177823	176044	336033	248387	172564	422947	407620	999775
Hsa-Mir-27-P3_3p	11134	4538	6316	11208	8880	7110	6048	10429	11564	4314	3946	7213	5353	7333	8045	4711	4313	7001	7578	8606	4270	23050	18861
Hsa-Mir-28-P1_5p	1438	962	1189	3235	2113	1522	1002	2937	2762	713	798	1705	1285	1138	1421	826	943	1459	1316	1527	894	1703	3739
Hsa-Mir-28-P1_3p	14574	10434	9908	25120	17403	12641	8493	20984	22652	5836	5997	12424	12393	12443	12565	4986	7353	12725	11128	9062	8432	19193	36351
Hsa-Mir-28-P2_5p	29328	38256	69875	55300	108618	47514	51180	55486	74781	29302	26416	44621	34429	37379	72979	34109	26047	51962	42063	38068	57885	33314	153639
Hsa-Mir-28-P2_3p	6468	15234	25710	14939	35513	11334	17850	19264	18176	9379	6398	13050	9831	11713	24988	6739	9175	15407	12966	7155	20470	46720	45029
Hsa-Mir-28-P3_5p	1337	1483	1943	1113	1909	763	1537	2643	2515	1419	1089	1245	1553	1502	1909	1048	848	1801	1498	2654	1302	8424	5772
Hsa-Mir-29-P1b-v1_3p	6548	9233	9231	7204	18075	4033	10383	18227	16612	7804	3548	6990	9338	12501	9461	5152	6062	8918	11360	7076	3688	35669	28289
Hsa-Mir-29-P1b-v2_3p	2989	3091	3800	3802	5576	2351	3578	7360	6025	2854	1926	2931	3390	4447	3581	2432	2387	2846	4032	4117	2158	11189	10093
Hsa-Mir-29-P1d-v1_3p	6548	9233	9231	7204	18075	4033	10383	18227	16612	7804	3548	6990	9338	12501	9461	5152	6062	8918	11360	7076	3688	35669	28289
Hsa-Mir-29-P1d-v2_3p	2989	3091	3800	3802	5576	2351	3578	7360	6025	2854	1926	2931	3390	4447	3581	2432	2387	2846	4032	4117	2158	11189	10093
Hsa-Mir-29-P2b_3p	43894	40611	47851	60529	67338	32490	40318	87923	86030	31781	24682	40846	39369	43362	43606	30538	27862	41001	52719	64033	26618	171798	127958
Hsa-Mir-29-P2d_3p	11586	13395	16570	19337	23043	9869	13317	28764	27061	10062	7965	14105	14032	14217	14330	9693	9690	13547	16568	18645	8055	57777	41609
Hsa-Mir-30-P1a_5p	79701	112919	97895	141169	120517	88055	76955	157223	225215	57601	63546	110045	110091	100389	115458	56006	65757	124821	105362	101044	77062	349596	403791
Hsa-Mir-30-P1b_5p	187929	181079	154770	275225	282422	133689	153308	400714	388430	128120	98963	196102	201672	262121	206834	76514	156644	202675	219067	139165	101046	563598	585382
Hsa-Mir-30-P1d_5p	81149	77627	94091	119188	152861	69828	75424	156264	183906	64357	51187	96291	103617	103390	104392	53593	67159	95959	104021	77035	55954	335059	311134
Hsa-Mir-30-P2a_5p	14185	20833	19681	21785	22050	12162	16435	37164	40028	12170	10449	20531	20408	22148	21349	9002	13257	18881	22854	19332	13633	32911	71196
Hsa-Mir-30-P2b_5p	36070	38564	46605	79221	61147	44997	34395	89158	95562	23331	24714	50059	51940	48346	45494	23380	32108	54920	47955	44324	26101	69221	161776
Hsa-Mir-30-P2d_5p	36070	38564	46605	79221	61147	44997	34395	89158	95562	23331	24714	50059	51940	48346	45494	23380	32108	54920	47955	44324	26101	69221	161776
Hsa-Mir-31_5p	949	1215	1564	2112	1607	1396	1339	2686	3318	1531	1199	1243	1547	1283	1492	1219	1063	1899	1369	1313	873	2883	6805
Hsa-Mir-32_5p	293	336	824	604	1123	324	539	663	651	344	205	450	333	575	895	305	247	431	356	267	354	491	1286
Hsa-Mir-33-P1_5p	433	1173	2449	1384	4399	927	1858	985	1114	737	446	775	350	793	2830	949	653	1175	893	337	1031	1586	3451
Hsa-Mir-33-P3_5p	323	523	616	493	1112	253	521	462	632	298	179	310	209	458	787	263	294	494	481	332	166	3812	1312
Hsa-Mir-34-P1_5p	967	1588	1556	1645	2428	1377	1840	2643	4042	1174	910	1797	1995	1732	1737	1376	924	2212	1860	1222	683	4965	7548
Hsa-Mir-34-P2a_5p	79	102	253	82	238	73	206	145	102	90	49	106	73	77	373	61	6136	58	52	177	74	408
Hsa-Mir-34-P2a_3p	17	15	56	16	34	21	44	14	34	16	10	18	12	15	54	13	739	9	17	56	21	84
Hsa-Mir-34-P2b_5p	589	992	1597	611	1228	475	1403	678	751	520	417	807	405	310	2047	509	305	1064	275	390	1336	802	2460
Hsa-Mir-34-P3a-v1_5p	7	11	6	5	16	12	8	17	13	14	6	11	11	13	5	7	811	20
Hsa-Mir-34-P3a-v2_5p	8	20	14	23	19	24	12	31	28	26	15	18	25	16	6	10	221	20	6	14	34
Hsa-Mir-34-P3b_5p	7	2	2	10	16	7	25	5	7	7	11	7	10	7	3	1	912	2	11	14
Hsa-Mir-34-P3b_3p	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	00