Quanta sybr green supermix

Relative gene expression was determined using a two-step quantitative real-time PCR method. Total RNA was isolated with the RNeasy Isolation kit with on-column DNase I treatment (Qiagen) and reverse-transcribed using the cDNA Synthesis Kit (Quanta biosciences). Quantitative reverse transcription PCR (RT–PCR) was performed with the Quanta SYBR Green Supermix (Quanta biosciences) on the ABI Prism 7500 Real-Time PCR System (Applied Biosystems). Fold changes in gene expression were determined using the comparative C_T method (ΔΔCt) with normalization to the housekeeping genes GAPDH or B2M. The primer sequences used in the study are shown in Supplementary Table 2. The TaqMan hPSC Scorecard panel was used to assess pluripotency and trilineage differentiation potential according to manufacturer's instructions (Life Technologies). Data analysis was performed using the hPSC Scorecard software (Life Technologies).

Relative gene expression was determined using a two-steps quantitative real-time PCR method. Total RNA was isolated with the RNeasy Isolation kit with on-column DNase I treatment (Qiagen, Germany) and reverse-transcribed using the cDNA Synthesis Kit (Biorad, Hercules, CA). Quantitative RT-PCR was performed with the Quanta SYBR Green Supermix (Quanta biosciences, Beverly, MA) on the ABI Prism 7500 Real Time PCR System (Applied Biosystems, Foster City, CA). Fold changes in gene expression were determined using the comparative CT method (△△Ct) with normalization to the housekeeping gene B2M.