GSM8478649: t-BHP 3; Candida albicans; RNA-Seq
RNA was isolated from a frozen cell pellet containing 1x10^8 cells using the RNeasy Plus Universal Mini Kit, following the manufacturer's guidelines (Qiagen, Germantown, MD). The RNA concentration was determined using a Qubit 2.0 Fluorometer (Life Technologies), and RNA integrity was checked with an Agilent TapeStation 4200 (Agilent Technologies, Santa Clara, CA). For sequencing, RNA samples were processed using the NEBNext Ultra II RNA Library Prep Kit for Illumina, adhering to the manufacturer's protocol (NEB, Ipswich, MA). Briefly, mRNA was enriched with Oligo(dT) beads, fragmented at 94°C for 15 m, and used to synthesize cDNA. The cDNA fragments were end-repaired, adenylated at the 3' ends, and ligated with universal adapters, followed by index addition and library amplification via limited-cycle PCR. The sequencing libraries were validated using an Agilent TapeStation (Agilent Technologies), quantified using a Qubit 2.0 Fluorometer (Invitrogen), and assessed by quantitative PCR (KAPA Biosystems). The libraries were pooled, clustered on a single lane of a flowcell, and sequenced on an Illumina HiSeq instrument (4000 or equivalent) using a 2x150 bp Paired End (PE) configuration.