Targeted DNA sequencing of biochemically enriched regions of a yeast genome
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DNA-binding proteins are key factors involved in the maintenance of genome integrity and gene expression. To identify loci that are recognized by a Myb domain containing DNA-binding protein Tay1 in the genomic DNA of the yeast Yarrowia lipolytica [1, 2], we combined a pulldown assay and DNA sequencing on a MinION device. The sequencing library has been prepared using 1D ligation chemistry (SQK-LSK108) and sequenced in a FLO-MIN106 (R9.4.1) flow cell. Resulting sequence data were mapped to the reference genome. The comparison with the data obtained by sequencing of a control DNA sample revealed a number of genomic DNA fragments including the telomeric sequences that were biochemically enriched in the pull-down experiment. This approach can be useful in genome-wide identification of loci potentially recognized by DNA-binding proteins of interest as well characterization of sequences representing their putative binding sites.
