Superscript 3 qrt pcr kit

Cells were harvested and total RNA was isolated using TRIzol reagent (Invitrogen). A SuperScript III qRT-PCR kit (Invitrogen) was used to synthesize cDNA from total RNA. Quantitative PCR was performed using a ViiA7 system (Applied Biosystems, Rockford, IL) or LightCycler 480 system (Roche, Indianapolis, IN) with iTaq Universal SYBR Green Master Mix (BioRad); conditions were 95 °C for 10 min followed by 50 cycles at 95 °C for 15 s and 60 °C for 3 s. Samples were run in triplicate and transcripts were quantitated by comparing Cycle Threshold (Ct) values for each reaction with a GAPDH reference. Primer sets for quantitative PCR are listed in Supplementary Table 3.

Cells were harvested and total RNA was isolated using TRIzol reagent (Invitrogen). A SuperScript III qRT-PCR Kit (Invitrogen) was used to synthesize cDNA from total RNA. qPCR was performed using a ViiA7 system (Applied Biosystems) with iTaq Universal SYBR Green Master Mix (Bio-Rad); conditions were 95 °C for 10 min followed by 50 cycles at 95 °C for 15 s and 60 °C for 3 s. Samples were run in triplicate and Klf4, Stau1, Nestin, NeuN, Dlx1, Dlx2, Tuj1, and Gad67 transcripts were quantitated by comparing cycle threshold (Ct) values for each reaction with a Gapdh reference. Primer sets for quantitative PCR are listed in Supplementary Table 2.

Cells were harvested and total RNA was isolated using TRIzol reagent (Invitrogen). The SuperScript III qRT-PCR kit (Invitrogen) was used to synthesize cDNA from total RNA. Quantitative PCR was carried out using the ABI PRISM 7900 Sequence Detection System with SYBR Green Master Mix (iTaq) with conditions of 95°C for 10 min followed by 50 cycles at 95°C for 15 sec and 60°C for 3 sec. Samples were run in triplicate and Dlx1, Dlx2, Tlx3, NeuroD1, Tuj1, Gad67, NeuN, Mbp, Gfap, Ascl1, and Id1 transcript quantitation was undertaken by comparing Cycle Threshold (Ct) values for each reaction with the Gapdh reference. Primer sets for quantitative PCR are listed in S5 Table.