Whole genome sequencing of Candida albicans SC5314 and SC5314-derived BWP17 SIR2 deletion mutants
Whole genome sequencing of Candida albicans strain BWP17 (a derivative of the common laboratory strain SC5314) and three SIR2 deletion mutants built from it (one MTLa/alpha, one MTLa/a, one MTLalpha/alpha). The primary purpose was to genotype the SIR2 deletion mutants in order to identify genetic variations that could be contributing to observed altered phenotypes. Briefly, -80C frozen stocks were struck to solid YPD medium and incubated at 30C for 2 days. Cells from the densest part of the streaks were then used to start overnight cultures in liquid YPD medium at 30C. The next day, ~10^8 cells were used to extract genomic DNA with the Zymogen Quick-DNA Fungal/Bacterial Miniprep Kit. The constructed libraries were dual-barcoded and sequenced as 2x150 paired-end reads on an Illumina NextSeq 2000. After sequencing, the reads were demultiplexed and Illumina adapters were trimmed. Sequencing was also performed on a CRISPR-competent SC5314 strain, with methods as described above. Additional sequencing of CRISPR-competent SC5314 SIR2 mutants and SIR2-complemented mutants in both backgrounds was performed. The primary purpose was to genotype additional SIR2 deletion mutants in order to identify genetic variations that could be contributing to observed altered phenotypes. Strains were struck onto solid YPD medium and grown at 30C for two days, after which the strains were processed for sequencing by SeqCoast Genomics (Portsmouth, NH, USA). Briefly, DNA from each strain was extracted using the MagMAX Microbiome Ultra Nucleic Acid Isolation Kit and prepared for whole genome sequencing using the Illumina DNA Prep tagmentation kit and unique dual indexes. Sequencing was performed on the Illumina NextSeq2000 platform to produce 2x150bp paired reads. Reads were demultiplexed, trimmed, and assessed using FastQC.