The LightCycler® 480 real-time PCR system: a versatile platform for genetic variation research

Genetic control of biennial bearing in apple

“…After activation at 95 °C for 5 min, the reactions underwent 40 PCR cycles of: 95 °C for 10 s; 55 °C for 30 s; 72 °C for 15 s. The HRM analysis was performed immediately after the PCR amplification, with single steps at 95 °C for 1 min; 40 °C for 1 min; 65 °C for 1 s; and then a slow increase of the temperature to reach 95 °C over 15 min, with continuous measurement of the fluorescence intensity (25 data points per degree Celsius). The Roche software identified sequence variants as groups that exhibit similar melting profiles ( Hoffmann et al , 2008 ).…”

RNAi-Mediated Silencing of Laccase 2 in Culex pipiens Pupae via Dehydration and Soaking Results in Multiple Defects in Cuticular Development

“…No template and no RT controls were included in each reaction. qRT-PCR was performed on a Roche Lightcycler 480 System [33] using SYBR Green master mix (Bio-Rad Laboratories, Inc. Hercules, CA, USA) and laccase 2-specific primers (Table 1). The reactions were performed for 1 min at 95 • C, followed by 39 cycles of 10 s at 95 • C, 30 s at 58 • C, and 30 s at 72 • C. The melting curve was analyzed at 60-95 • C to ensure a single peak, and thereby primer specificity.…”