Total RNA was isolated using a TRIzol isolation kit. Whole cDNA was synthesized from 1 µg of purified RNA by the SuperScript II First-Strand cDNA synthesis system (Invitrogen). Primer sequences for p62 qRT-PCR were described previously by Hu et al. (2012) (link). The qRT-PCR was performed with SYBR green brilliant III ultrafast reagent kit (MX3000P; Agilent Technologies) following the manufacturer’s protocol. The results of qRT-PCR assays presented are a mean of three independent RNA preparations. Each sample was analyzed in triplicates.
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qRT-PCR Analysis of p62 Expression
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Corresponding Organization : Emory University
Other organizations : Southeast University
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Variable analysis
independent variables
- None explicitly mentioned
- Expression level of p62 gene (as measured by qRT-PCR)
- Purified RNA input amount (1 μg)
- Reverse transcription protocol (SuperScript II First-Strand cDNA synthesis system)
- QRT-PCR protocol (SYBR green brilliant III ultrafast reagent kit, MX3000P; Agilent Technologies)
- Number of independent RNA preparations (three)
- Number of technical replicates per sample (triplicates)
- No positive or negative controls were explicitly mentioned in the information provided.
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