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. 2019 Sep 5;14(9):e0222170. doi: 10.1371/journal.pone.0222170

Fig 1. Amplification curves of thermal gradient RT-LAMP for optimizing annealing temperature for Citrus tristeza virus (CTV) VT isolate specific LAMP primers.

Fig 1

(a) RT-LAMP reactions with an annealing temperature gradient ranged from 59°C to 66°C was set using the Genie III. CA-VT-AT39 RNA detection times were 11:00, 10:35, 9:40, 9:00, 8:35, 8:20, 8:15, and 8:25 (min:sec) at temperatures 59°C, 60°C, 61°C, 62°C, 63°C, 64°C, 65°C, 66°C, and 67°C, respectively. (b) Melting curve of thermal gradient RT-LAMP primers of Citrus tristeza virus CA-VT-AT39 isolate after amplification for 20 min and detected on the FAM channel using 1°C steps, and a hold of 30 sec at each step from 59 to 96°C. The viral RNA had a melting temperature (Tm) of 88.4±0.2°C indicating similar sequences and amplicon size.