Computational protocol: Targeted Metabolomics of Serum Acylcarnitines Evaluates Hepatoprotective Effect of Wuzhi Tablet (Schisandra sphenanthera Extract) against Acute Acetaminophen Toxicity

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Protocol publication

[…] Total RNA was extracted from frozen liver samples with Trizol reagent and quantified with NanoDrop (Thermo Scientific, Rockford, IL). cDNA was synthesized from 1 μg total RNA using Superscript II reverse transcriptase kit (Invitrogen, Carlsbad, CA). For quantitative real-time PCR (qPCR) analysis, the primer pairs were designed using the Primer Express software (Applied Biosystems) and all primers were further validated using NCBI Primer-BLAST. The primer sequences have been published []. qPCR reactions contained 25 ng of cDNA, 150 nM of each primer, and 5 μL of SYBR Green PCR Master Mix (Applied Biosystems, Foster City, CA) in a total volume of 10 μL. All reactions were performed in triplicate on an Applied Biosystems Prism 7900HT Sequence Detection System (Applied Biosystems, Foster City, CA). Relative mRNA levels were calculated by the comparative threshold cycle method using β-actin as the internal control. […]

Pipeline specifications

Software tools Primer Express, Primer-BLAST
Application qPCR
Diseases Liver Diseases, Chemical and Drug Induced Liver Injury, Drug-Related Side Effects and Adverse Reactions
Chemicals Acetaminophen