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. 2018 Jan 3;13(1):e0190523. doi: 10.1371/journal.pone.0190523

Fig 2. pot1Δ pic1-T269 double mutants lose telomeric DNA and have circular chromosomes.

Fig 2

(A) The telomere length of pot1Δ pic1-T269 double mutants was analyzed by Southern hybridization at 25°C. pot1Δ and pic1-T269 single mutants were used as a control for strains that lost and retained the telomeric DNA, respectively. Genomic DNA was digested by EcoRI and fractionated by 1.5% agarose gel electrophoresis. Telomere plus telomere associated sequence (TSA1) derived from pNSU70 was used as a probe for hybridization. To assess the total amount of DNA, the gel was stained with ethidium bromide (EtBr) before blotting onto the membrane. (B) Restriction enzyme sites around the telomere and TAS1 of one chromosome arm cloned in the plasmid pNSU70. (C) NotI-digested chromosomal DNA from pic1-T269, pot1Δ and pot1Δ pic1-T269 cells were analyzed by PFGE at 25°C. The digested DNA fractionated in a 1% agarose gel and a mixture of four probes (L, I, C, M) specific to the NotI-digested chromosomal terminal fragments were used. (D) NotI restriction enzyme map of S. pombe chromosomes.