Total rna extraction kit

Total ribonucleic acid (RNA) was extracted and purified from freshly isolated rat renal cortical tissues using a total RNA extraction kit (Amresco, Columbus, OH, USA). Briefly, rat renal cortical tissue was homogenized in extraction reagent as per the manufacturer’s protocol. The purity and integrity of total RNA were determined by a multidetection microplate reader (Biotek, Winooski, VT, USA). Subsequently, first-strand complementary deoxyribonucleic acid (cDNA) was synthesized using the iScript™ cDNA synthesis kit (Bio-Rad, Hercules, CA, USA). mRNA levels were determined quantitatively using the Sensi-FAST SYBR Lo-Rox Kit (Bioline, London, UK). Primer sets were used as previously published [13 (link)] and were purchased from Integrated DNA Technologies (Coralville, IA, USA). Gene expressions were normalized to the β-actin mRNA level and expressed as relative fold changes (RFC). qPCR amplification was performed in duplicate for each cDNA.

Total RNA was purified from freshly isolated rat renal cortical tissues using total RNA extraction kit (Amresco, OH, USA), according to the manufacturer's instruction. The first strand cDNA was obtained using iScript cDNA synthesis kit (Bio-Rad, CA, USA) and qPCR was performed using SYBR real-time PCR master mix (Toyobo, Osaka, Japan) on ABI 7500 (Life Technologies, NY, USA). Primers were designed according to published sequences (Table 1) and were purchased from Biobasic Inc. (ON, Canada). Gene expressions were normalized to actin mRNA levels and reported as relative fold changes (RFC). QPCR amplification was performed in duplicate for each cDNA.

Total RNA was extracted from hippocampi using a total RNA extraction kit (Ribozol, Amresco) following manufacturer’s instructions. 5 μg of RNA in 10 μl was used for cDNA synthesis. First-strand cDNA was synthesized by RT using oligo (dT) primers and SuperScript II reverse transcriptase (Invitrogen). For the PCR array, total RNA (100 ng) was amplified using SYBR Green Master Mix (ThermoFisher) and analyzed using a CFX96 qPCR instrument (Bio-rad).