Figure 5.

Analysis of the isochromosome centromere. (A) Schematic of predicted centromere structure of Cen-Ch¹⁶ based on homologous cen3, with the break point of Ch¹⁶-RMGAH and Ch¹⁶-YAMGH-derived LOH colonies, indicated by an arrow. cnt3, innermost (imr3), outer (otr3) and irc3 repeats are shown. (B) Log₂ ratio of the fold coverage from Illumina GAII sequencing across isolated isochromosome and minichromosome DNA (log₂ of isochromosome/minichromosome) from strains TH4313 and TH2125, respectively. Sequence data aligned to S. pombe ChIII reference sequence at positions indicated (C) PCR amplification of irc3L and irc3R using irc3-R and irc3-F primers, digested using ApoI, (D) PCR amplification of imr3–otr3 junctions using imr-out and dh primers (E) PCR amplification of imr3L–cnt3 junction using primers cnt3-L and imr3-in (F) PCR amplification of cnt3–imr3R junction using primers cnt3-R and imr3-in, separated on a 2% agarose gel and stained with EtBr. Diagnostic PCR product sizes are shown. PCR amplification was carried out as described previously (Nakamura et al, 2008).