Skip to main content
. 2018 Mar 23;13(3):e0194412. doi: 10.1371/journal.pone.0194412

Table 1. Standard reaction mixes and cycling conditions.

Mastermix Preparation Cycling Conditions
Superscript-III kit
SuperScript™ III One-Step RT-PCR System with Platinum™ Taq DNA Polymerase		 Total Reaction volume 25 μL:
1) x2 Mastermix 12.5 μL
2) Forward and reverse primers 400nM
3) Probe 160nM
4) Enzyme 0.5 μL
5) JEV RNA 5 μL		 50°C for 15 minutes
95°C for 2 minutes
X 45 (95°C for 15 seconds + 60°C for 45 seconds)
Express kit
EXPRESS One-Step SuperScript® qRT-PCR		 Total Reaction volume 20 μL:
1) x2 Mastermix 10 μL
2) Forward and reverse primers 400nM
3) Probe 160nM
4) Enzyme 2 μL
5) JEV RNA 5 μL		 50°C for 15 minutes
95°C for 2 minutes
45 x (95°C for 15 seconds + 60°C for 45 seconds)
Fastvirus kit
TaqMan® Fast Virus 1-Step		 Total Reaction volume 25 μL:
1) X4 Mastermix 6.25 μL
2) Forward and reverse primers 400nM
3) Probe 160nM
4) JEV RNA 5 μL		 50°C for 5 minutes
95°C for 20 seconds
45 x (95°C for 15 seconds + 60°C for 60 seconds)