Quantitect retrotranscription kit

Total RNA isolation was performed using a NucleoSpin RNA XS kit (Macherey-Nagel), according to the manufacturer’s instructions. RNA concentration and purity were determined using a NanoDrop ND-1000 Spectrophotometer (Thermo Fisher Scientific).
Reverse transcription was carried out with 250 ng of total RNA using the QuantiTect retrotranscription kit (Qiagen). Quantitative PCR (qPCR) reactions were performed using Taqman Array Fast plates and Taqman Gene expression master mix (Thermo Fisher Scientific) in an Applied Biosystems real-time PCR machine (7500 Fast System). All samples were normalized against a housekeeping gene (18S) and the gene expression was determined based on the ΔΔCT method. Average values were obtained from at least four biological replicates. The primer sets and MGB probes (Thermo Fisher Scientific) labelled with FAM for amplification are listed in Supplementary Table 3.

Total RNA isolation was performed using a RNeasy Mini Kit (Qiagen), according to the manufacturer’s instructions. RNA concentration and purity were determined using a NanoDrop ND-1000 Spectrophotometer (Thermo Fisher Scientific).
Reverse transcription was carried out with 250 ng of total RNA using the QuantiTect retrotranscription kit (Qiagen). Quantitative PCR (qPCR) reactions were performed using Taqman Array Fast plates and Taqman Gene expression master mix (Thermo Fisher Scientific) for CRX and BRN3B and S18 in an Applied Biosystems real-time PCR machine (7500 Fast System). All samples were normalized against a housekeeping gene (18S) and the gene expression was determined based on the ΔΔCT method relative to D35. Average values were obtained from at least four biological replicates. The primer sets and MGB probes (Thermo Fisher Scientific) labeled with FAM for amplification are listed in Table 3.

Total RNA was extracted using a RNeasy mini kit (Qiagen, Düsseldorf, Germany) following the manufacturer’s instructions. RNA concentration and purity were determined using a NanoDrop ND-1000 Spectrophotometer (Thermo Scientific). Reverse transcription was carried out with 1 μg of total RNA using the QuantiTect retrotranscription kit (Qiagen). All the samples were normalized against a housekeeping gene (GAPDH). The primer pairs (IDT) are listed in Table 3 All RT-PCR reactions were run for 40 cycles of 94 °C for 20 s, 58 °C for 1 min and 68 °C for 30 s.