H3K4me3 ChIP-seq of Candida albicans WT: IP replicate 1
20 OD600 units of exponential phase cells were fixed with 1% formaldehyde for 15 minutes at room temperature. Reactions were quenched by the addition of glycine to a final concentration of 125 mM. Cells were lysed using acid-washed glass beads and a DisrurGenie for four cycles of 30 minutes at 4C with 5 minutes on ice between cycles. Chromatin was sheared to 200500 bp using a BioRuptor sonicator for a total of 20 minutes (30 seconds on, 30 seconds off cycle) at 4C. Immunoprecipitation was performed overnight at 4C using 2 l of the appropriate antibody and 25 l of protein G magnetic Dynabeads. ChIP DNA was eluted, and cross-links reversed at 65C in the presence of 1% SDS. All samples were then treated with RNaseA and proteinase K before being purified by phenol:chloroform extraction and ethanol precipitation. Libraries were prepared and sequenced as 50bp single end reads on an Illumina HiSeq2000 platform.