Transcriptomic Analysis of the Candida albicans Response to Treatment with Baicalin
In order to investigate differential gene expression at high resolution, here, we have performed RNA sequencing analysis of samples obtained from Candida albicans cells grown in the presence or absence of baicalin. Overall treatment with baicalin resulted in 496 up-regulated and 346 down-regulated genes. Overall design: Candida albicans strain SC5314 was grown in YPD medium overnight, washed with PBS and cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum at 2.5×10^4 CFU/mL and incubated at 37 ? for 24 hours in the presence or absence of baicalin at 500 µg/mL. Total RNA was extracted and mRNA enriched by magnetic beads with oligo(dT). The mRNA was randomly broken by adding fragmentation buffer. Short fragments of double-stranded cDNA generated by reverse transcription were subjected to end-repair, phosphorylation and 'A' base addition according to Illumina's library construction protocol. Libraries were size selected for cDNA target fragments of 300 bp on 2% Low Range Ultra Agarose followed by PCR amplified using Phusion DNA polymerase for 15 PCR cycles. After quantified by Qubit 4.0, paired-end RNA-seq sequencing library was sequenced with the NovaSeq 6000 sequencer (2 × 150 bp read length). Three biological replicates were obtained for each condition (treated and untreated).