The Ebola virus is spread through bodily fluids including saliva and sweat, and those who had worked in Sierra Leone were operating under the same strict "no touch" protocols as the public under the state emergency laws.
By being underfoot, the glasses were divorced from their function and protected only by the don't-touch protocol of museums.
Mission: Impossible Ghost Protocol?
This second PCR reaction consisted of 3 min at 94°C followed by a touch down protocol decreasing the annealing temperature from 60°C to 45°C during 40 s (−1°C/cycle), with 1 min extension and 40 s denaturalization steps at 72°C and 94°C, respectively, followed by 24 cycles of 40 s at 45°C, 72°C and 94°C, and a final elongation step of 7 min at 72°C.
After an initial denaturation (2 min at 94°C), a touch down protocol was used as follows: 10 cycles with a decreased hybridisation temperature from 61 to 58°C every two cycles.
A touch down protocol was adopted as follows [ 25]: After denaturation at 95°C for 5 min, the annealing temperature was decreased 0.5°C every cycle from 63°C to 56°C, at which temperature 35 cycles were carried out.
For PCR amplification of each exon, a touch down protocol was used as follows: initial denaturation and HotStar Taq Polymerase activation at 95°C for 15 min; six cycles of 30 s at 95°C, 30 s at 66°C (the annealing temperature decreasing by 2°C at every two cycles), 30 s at 72°C; followed by 40 cycles of 30 s at 95°C, 30 s at 60°C and 30 s at 72°C.
Cycling conditions followed the touch-down protocol of [12].
For TNF-α, the annealing conditions were modified to include a 63°C 59°C touch-down protocol, decreasing temperature by 0.5°C per cycle for the first 8 cycles.
Sequencing of one candidate (FG543) was correctly achieved for A. ostoyae although three had resulted in PCR success, each with one single product amplified with the PCR touch-down protocol.
