RNA-seq of human airway organoids and PAO1 pqsC
Total RNAs (2 g) were used for preparation of a stranded RNA sequencing library, using a KC-DigitalTM Stranded mRNA Library Prep Kit for Illumina (# DR08502, Wuhan Seqhealth, China), following the manufacturers instruction. The kit eliminated duplication bias in the PCR and sequencing steps, by using a unique molecular identifier (UMI) of eight random bases to label the pre-amplified cDNA molecules. The library products corresponding to 200500 bp were enriched, quantified, and finally sequenced using a Novaseq 6000 sequencer (Illumina) with a PE150 model.