Identification of centromeres in Candida tropicalis
– Libraries for multiplex ChIP Sequencing were constructed using NEXTflexTM ChIP-SeqSample Preparation Kitprotocol outlined in “Preparing Samples for ChIP Sequencing of DNA” (BIOO Scientific# IP-5143-01). Briefly, DNA was subjected to a series of enzymatic reactions that repair frayed ends, phosphorylate the fragments. The end repaired  fragments are adenylated with a single nucleotide „A‟ overhang (BIOO Scientific #IP-5143-01) and ligate adaptors (NEXT Flex adapters). The fragments with ligated adapters are subjected to pre  size selection PCR for 5 cycles followed by size selection on 2% Low Melting Agarose. The size selected samples are enriched with 13 cycles of PCR. The prepared libraries were quantified using Nanodrop Spectrophotometer and validated for quality by running an aliquot on High Sensitivity Bioanalyzer Chip (Agilent).