Total RNA was extracted from tissues and cells using TRIzol (Invitrogen, Thermo Fisher Scientific, Inc.). Then RNA (1 μg) was converted into cDNA using a SuperScript® One-Step RT-PCR kit (Thermo Fisher Scientific, Inc.), and qPCR was performed to examine the mRNA expression using a SuperScript® One-Step RT-PCR kit (Thermo Fisher Scientific, Inc.), according to the manufacturer’s protocol. The PCR reaction conditions were as follows: 95°C for 3 min, followed by 40 cycles at 95°C for 15 s and 60°C for 30 s. The relative expression levels were analyzed using the 2−ΔΔCq method21 (link).
Protocol detail
Quantitative Gene Expression Analysis
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Corresponding Organization :
Other organizations : 175th Hospital of People's Liberation Army, The 180th Hospital of PLA
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Variable analysis
independent variables
- None explicitly mentioned
- MRNA expression
- None explicitly mentioned
- Positive control: None mentioned
- Negative control: None mentioned
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