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. 2006 May 31;34(10):2914–2924. doi: 10.1093/nar/gkl293

Figure 3.

Figure 3

Analysis of split-chromosomes in the fob1 mutants by CHEF gel electrophoresis and Southern hybridization (A–C). Chromosomal DNA was isolated from the following strains: (A) ID-11, (B) ID-12 and (C) ID-13. (D) Analysis of rDNA copy numbers. Genomic DNA was digested at a unique KpnI site in the rDNA unit and subjected to electrophoresis followed by Southern hybridization using 5S rDNA as a probe. A single copy gene was used as an internal control for normalization. Because the normalization factor for the 5S rDNA probe was found to be 15 relative to the single copy gene, rDNA copy numbers were determined by multiplying by 15. (E) Relative quantification of 18S rRNA using real-time RT–PCR. The 18S rRNA amount is divided by the ACT1 amount to obtain a normalized 18S rRNA value and the normalized amount of 18S rRNA in FY833 (WT) was used to compare the relative amount of 18S rRNA in different strains.