Total transcriptome cdna synthesis kit

The mRNA levels of runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), collagen type I (COL-1), and bone morphogenetic protein 2 (BMP2) were assessed by qRT-PCR. The cells were first washed three times with cold PBS, and total RNA was isolated using the RNA Extraction Kit (Accurate Biology, AG21017). cDNA was synthesized using the Total Transcriptome CDNA Synthesis Kit (Accurate Biology, AG11707). cDNAs were normalized to GAPDH, respectively. Primers were designed with Primer Premier 5.0 software and listed in Table 1. A comparison of the 2^−ΔΔCt method was used to quantify the relative expression levels of mRNAs.

Samples of total RNA were reverse transcribed into cDNA using a Total-Transcriptome cDNA Synthesis Kit (AG11707, Accurate Biology, China) according to the manufacturer’s instructions. Quantitative real-time polymerase chain reaction measurements were performed using the StepOnePlus fluorescence quantitative PCR instrument (Applied Biosystems, Foster City, CA, United States) and a SYBR Green qPCR Mix according to the manufacturer’s instructions. The data were analyzed using the 2^–ΔΔCT method (Livak and Schmittgen, 2001 (link)), and the results were represented as relative expression levels, Actb (encoding actin) was the reference gene. The information on the primers used is shown in Table 1.