Ez 10 total rna mini preps kit reagent

Total RNA samples were extracted from the right hippocampus tissues using EZ-10 Total RNA Mini-Preps Kit Reagent (Sangon Biotech, Shanghai, China). The concentration and purity of total RNA were detected by Ultraviolet spectrophotometry (Jinghua, Shanghai, China). The synthesis of the first-strand cDNA was performed by the First Strand cDNA Synthesis Kit, according to the protocol of the manufacturer (ABclonal, Wuhan, China). The real-time PCR system was used for the detection of the quantitative RT-PCR (Bio-Rad, Shanghai, China). The 2^−ΔΔCt method was used for the relative expression level of each gene based on the reference expression of β-actin. The primer sequences used for PCR were as follows: parkin forward primer: 5′-GAGGTCGATTCTGACACCAGC-3'; parkin reverse primer: 5′-CCGGCAAAAA TCACACGCAG-3'; pink1 forward primer: 5′-TTCTTCCGCCAGTCGGTAG-3'; pink1 reverse primer: 5′-CTGCTTCTCCTCGATCAGCC-3'; LC3II forward primer: 5′-TTATAGAGCGATACAAGGGGGAG-3'; LC3II reverse primer: 5′-CGCCGTC TGATTATCTTGATGAG-3'; p62 forward primer: 5′-CATCGGAGGATCCGAGTG TG-3'; p62 reverse primer: 5′-TTCTTTTCCCTCCGTGCTCC-3'; beta-Actin forward primer: 5′-GGGAAAT CGTGCGTGACATTAAGG-3'; beta-Actin reverse primer: 5′-CAGGAAGGAAGGC TGGAAGAGTG-3'.