Sybr premix ex taq tli rnase plus

Total RNA was isolated from BV-2 cells using RNAiso Plus Reagent (Takara, Dalian, PR China), and reverse-transcribed by using PrimeScript^TMRT Reagent Kit (Takara, Dalian, China) to produce cDNA. The quantitative real-time PCR (qRT-PCR) was performed with SYBR^® Premix Ex Taq™ (Tli RNase Plus) (Takara, Dalian, china). Glyceraldehydes-3-phosphate dehydrogenase (GAPDH) mRNA was used as an internal control. The quantitative PCR program used was as follows: predenaturation (95°C, 5 min), denaturation (95°C, 20 sec), annealing (55°C, 20 sec), and extension (72°C, 45 sec), using primers specific for iNOS, COX-2, IL-6, IL‐1β and TNF-α.

Total RNA was isolated from rat MSCs under normoxic and hypoxic conditions using TRIzol Reagent, and cDNA was synthesized using a Maxime RT PreMix kit (iNtRON Biotechnology, Seongnam, Korea). The level of each gene transcript was determined quantitatively using a StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). A SYBR Green Dye system (SYBR Premix Ex Taq (Tli RNase Plus) with a ROX reference dye (TAKARA Bio Inc., Foster City, CA, USA) was used to perform real-time RT-PCR. All values are shown as the target gene expression level (fold change; 2ΔΔCt) normalized to the Gapdh transcript level. All primers were designed using Primer3 from BLAST (Table 1).

The level of each gene transcript was quantitatively determined using a StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). Total RNA was isolated from rat hearts using TRIzol reagent (Invitrogen), and reverse-transcription was performed using a Maxime RT Premix kit (iNtRON Biotechnology, Seongnam, Korea). A SYBR Green Dye system [SYBR Premix Ex Taq (Tli RNase Plus)] with an ROX reference dye (TaKaRa Bio Inc., Foster City, CA, USA) was used to perform real-time RT-PCR. The level of each gene transcript (FBXO11 and CREBZF) was normalized to GAPDH transcript levels, and relative changes in gene expression were quantified using the ∆∆CT method. Table 2 lists all the primers.