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Quantitative RT-PCR for casp-3, JNK, HO-1, and Keap-1 genes

EH Eman I. Hassanen
AH Ahmed M. Hussien
SM Sally Mehanna
MI Marwa A. Ibrahim
NH Neven H. Hassan
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The total RNA was extracted using the RNeasy Mini Kit (Qiagen Cat No./ID: 74,104) according to the instructions provided. The synthesis of the first-strand cDNA was performed using SuperScript Reverse Transcriptase (Thermo Scientific) according to the manufacturer’s instructions. Quantitative real-time PCR was done using SYBR™ Green PCR Master Mix (Thermo scientific Cat number: 4309155) by the ABI Prism Step One Plus Real-Time PCR System (Applied Biosystems). The assay was performed in duplicates and the ACTB was used as the internal standard for the calculation of the expression level. The primer sets of the studied genes were shown in Table ​Table11 and the fold change was calculated using 2−˄˄CT.

The primer sets of the studied genes

JNK, c-Jun N-terminal kinases; HO-1, Heme oxygenase; Keap-1, Kelch Like ECH Associated Protein 1; ACTB, Beta actin (housekeeping gene)

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