2.5. Colony PCR reactions
We used the Phusion® High-Fidelity PCR Kit for the Colony PCR and
followed the manufacturer’s protocols. We carried out the LAMP reactions
using the F3 and B3 primers. The medium-sized colonies (1.5 mm – 2 mm)
were selected from the plates and transferred into the reaction mixture.
Colony PCR was performed using a thermal cycler (Master cycler 384,
Eppendorf AG, Hamburg, Germany). Thermocycling conditions were set:
98 °C 30 s followed by 30 cycles of 98 °C 10 s, 56 °C and 72 °C 30 s,
and final extension was at 72 °C 10 min. The PCR products were observed
using 1.5 % agarose gel electrophoresis (Amplicon: 240 base pairs
(bp)).