Urine samples collection

Urine leftover samples collected on field, transferred to lab, aliquoted, and stored at -20°C.

Prepare urinary cf-urine and purification

Thaw urine, centrifuge, and extract miRNA using Norgen purification kit.

miRNA library preparation

Process miRNA library using Nextflex kit; check concentration and size with Qubit and TapeStation.

miRNA illumina NextSeq 550dx sequencing

Run library on Illumina NextSeq 550, 75 bp single-end flow cell; final concentration 1.8 pM.

Data analysis

Check raw reads quality with FastQC/multiQC; clean reads with Cutadapt; analyze with miRDeep2 and edgeR.

MM