Alui and mboi

Telomere gels were performed using telomere restriction fragment (TRF) analysis. Genomic DNA was digested using AluI and MboI (NEB). 4–10 μg of DNA was run on a 1% PFGE agarose gel (Bio-Rad) in 0.5× TBE buffer using the CHEF-DRII system (Bio-Rad) at 6 V cm⁻¹; initial switch time 5 s, final switch time 5 s, for 16 h at 14 °C. The gel was then dried for 4 h at 50 °C, denatured in a 0.5 N NaOH 1.5 M NaCl solution, and neutralized. Gel was hybridized with ³²P-labelled (CCCTAA)₆ oligonucleotides in Church buffer overnight at 42 °C. The next day, the membrane was washed four times in 4× SSC buffer, exposed onto a storage phosphor screen (GE Healthcare) and scanned using STORM 860 with ImageQuant (Molecular Dynamics). Telomere length was determined using TeloTool software^{35 (link)}.

Telomere gels were performed using telomere restriction fragment (TRF) analysis. Genomic DNA was digested using AluI and MboI (NEB). Then, 4–10 μg of DNA was run on a 1% PFGE agarose gel (Bio-Rad) in 0.5× TBE buffer using the CHEF-DRII system (Bio-Rad) at 6 V cm⁻¹; the initial switch time was 1 s, and the final switch time 6 s, for 17 h at 14 °C. The gel was then dried for 2 h at 60 °C, denatured in a 0.5 M NaOH/1.5 M NaCl solution, and neutralized. Gel was hybridized with ³²P-labeled (TTAGGG)₄ oligonucleotides in Church buffer overnight at 55 °C. The next day, the membrane was washed three times in 2× SSC buffer and once in 2× SSC 0.5% SDS, exposed onto a storage phosphor screen and scanned using Typhoon 9400 PhosphoImager (GE Healthcare). Telomere length was determined using TeloTool software.