The thermo cycling conditions chosen for PCR involved: a temperature ramp to 95 °C, 10 min hold; a reduction to 60 °C for 2 min; before increasing to 98 °C for 30 sec. This cycle between 60 °C and 98 °C was repeated 40 times to allow for efficient gene amplification. The system was then lowered to 60 °C and held for 2 min, before cooling to room temperature. After cooling, each chip was processed using the QuantStudio 3D Digital PCR system chip reader. To analyse chips, the AnalysisSuite™ software was used for quantification of the target gene. Each DNA sample was run in duplicate for each gene investigated.