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.