High-Throughput qPCR (HT-qPCR) chip (AsChip)

The diversity and abundance of ABGs in mouse gut microbiome were estimated using the Wafergen SmartChip Real-Time PCR System (WaferGen Biosystems, Fremont, CA). A total of 80 primer sets was designed to target 19 ABGs and one 16S rRNA gene. The reaction system (100 nL) for HT-qPCR including primers, LightCycler 480 SYBR Green I Master Mix, bovine serum albumin and DNA template. The thermal cycling conditions were 95 °C pre-denaturation for 10 min, 40 cycles of denaturation (95 °C, 30 s) and annealing (58 °C, 30 s, simultaneous annealing and extension).