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Denatured peptides look identical to intact peptides in solution you cannot visually detect the damage but they produce inconsistent or absent experimental outcomes

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To quantify mRNA, following cDNA synthesis, the quantitative RT-PCR (qRT-PCR) analysis was carried out on an Applied Rotor-Gene 3000 with the following conditions: GAPDH and target genes were amplified for 40 cycles at 94C for 30 seconds, 63C for 60 seconds, and 72C for 90 seconds