2.12 | Real-Time Reverse Transcriptase-PCR
Real-Time Reverse Transcriptase-PCR Total RNA was collected from kidney tissues and mTECs, using TRIzol reagents (Invitrogen). cDNA was synthesized using the Thermoscript RT-PCR synthesis kit (Fermentas, Pittsburgh, PA, USA) according to the manufacturer’s protocol. Real-time quantitative PCR analyses for mRNA were performed by using Thermoscript RT-qPCR kits (Fermentas, Pittsburgh, PA, USA) in an ABI PRISM StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). The products were used as templates for amplification using the SYBR Green PCR amplification reagent (Qiagen, Valencia, CA, USA) and gene-specific primers. Primers of genes in mice and mTECs were listed in Table S1.