High-resolution melting analyses of telomerase were performed using 1 L of end-point diluted fragmented DNA (~1 ng/L) coupled with the high-resolution melting reagent in a PCR in a total reaction volume of 3.5 L. High-resolution melting was performed on a Roche LightCycler 480 using standard settings (post-amplification: 10 L; high-resolution: 0.5 L; ramp rate: 0.1°C/s). All PCR components were prepared as proprietary cocktails: 1 L distilled water (final), 2.25 L HRM master mix (Roche Applied Systems), 0.5 L 4 M primer mix, 0.125 L 25 mM MgCl2. A short stage heating was run at 95°C for 5 minutes for reactions using SYBR Green I as dye. All reactions were performed in triplicate. Data analysis and plotting were performed as described in the manufacturer's manual. A representative plot can be found as Supplementary Material online at this link.
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