Renal RNAseq of WT during Candida albicans infection
Total RNA was isolated and RNA sequencing was performed by Novogene Corporation Inc. (Sacramento, USA). mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. To generate the cDNA library the first cDNA strand was synthesized using random hexamer primer and M-MuLV Reverse Transcriptase (RNase H-). Second strand cDNA synthesis was subsequently performed using DNA Polymerase I and RNase H. Double-stranded cDNA was purified using AMPure XP beads and remaining overhangs of the purified double-stranded cDNA were converted into blunt ends via exonuclease/polymerase. After 3' end adenylation a NEBNext Adaptor with hairpin loop structure was ligated to prepare for hybridization. In order to select cDNA fragments of 150~200 bp in length, the library fragments were purified with the AMPure XP system (Beckman Coulter, Beverly, USA). Finally, PCR amplification was performed and PCR products were purified using AMPure XP beads. The samples were read on an Illumina NovaSeq 6000 with 20 million read pair per sample.