GSM8988595: Ctrl-2; Candida albicans; RNA-Seq
For each sample, 1 mL of the culture was collected for RNA extraction using the Column Fungal RNA Extraction Kit (KANLANG, KL-10112BTN), following strict sterile and enzyme-free protocols. RNA integrity was assessed using the Agilent 4150 system, and only samples meeting quality thresholds were used for cDNA library construction. Sequencing was performed on the DNBSEQ-T7 (MGI) platform. After the sample passes quality control, eukaryotic mRNA is enriched using Oligo(dT)-attached magnetic beads. The mRNA is then randomly fragmented by adding a fragmentation buffer. Using the mRNA as a template, the first strand of cDNA is synthesized with random hexamer primers. Subsequently, the second strand of cDNA is synthesized by adding buffer, dNTPs, and DNA polymerase I. The double-stranded cDNA is purified using AMPure XP beads. The purified double-stranded cDNA then undergoes end repair, A-tailing, and adapter ligation, followed by size selection with AMPure XP beads. Finally, PCR amplification is performed to enrich the final cDNA library.