GSM8243296: Pool, YNB30, NO DOX, Set3, BioRep C; Candida albicans; OTHER
Genomic DNA was purified using the PureLinkTM Genomic DNA Mini Kit as per instructions with minor modifications, as described. Cell pellets were resuspended in 400 µL of lysis buffer (from PureLink Kit) and physically lysed by adding approximately 200 uL of acid-washed glass beads and bead beating six times for 30 seconds. Extracted genomic DNA was quantified with the Quant-iTTM PicoGreen dsDNA Assay Kit. Barcode amplification by PCR was performed using 105.6 ng of genomic DNA via the Ex-Taq enzyme UPTAG and DNTAG primer sequences are included in File S5 of associated paper. Separate UPTAG and DNTAG multiplexed pools were formed by combining equal amounts of PCR product from each sample. Pooled UPTAG and DNTAG DNA pools were extracted from a 1% agarose gel using the QIAquick® Gel Extraction Kit. Equal quantities of UPTAG and DNTAG pools as determined by PicoGreen assay were combined to form a library, which was sequenced on an Illumina MiSeq v2 Regular or NextSeq v2.5 Mid-Output instrument (single-end flow cell) using specific primers to sequence and index the UPTAG and DNTAGs (File S5).