Total RNA was isolated from cells using Sepasol-RNA I Super G RNA-isolation kit (Nacalai Tesque #09379). After the removal of DNA contamination by DNase I (Nippon Gene #312-05951), the total RNA was reverse-transcribed with ReverTra Ace qPCR RT Master Mix (TOYOBO #FSQ-201) to synthesize cDNA. Quantitative real-time PCR (qRT-PCR) was performed with THUNDERBIRD SYBR qPCR Mix (TOYOBO #QPS-201), using StepOnePlus and StepOne Software version 2.3 (Thermo fisher scientific). Sequences of the specific primer sets used in this PCR were listed in Supplementary Table 2 . The relative expression levels of the genes were calculated by ΔΔCt method normalized by Gapdh.
Sepasol rna 1 super g rna isolation kit
Manufactured by Nacalai Tesque
The Sepasol-RNA I Super G RNA-isolation kit is a product designed for the extraction and purification of RNA from various biological samples. It utilizes a guanidinium-based lysis and precipitation method to isolate high-quality RNA.
Automatically generated - may contain errors
2 protocols using sepasol rna 1 super g rna isolation kit
1
Quantitative Gene Expression Analysis
2
Histopathological Assessment of Intradermal OVA-Induced Inflammation
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For examination of local inflammation, mice were injected with OVA, OVA + MA or OVA + CFA intradermally at the ear. Injected sites in the ear were fixed and stained with hematoxylin and eosin for histopathological examination. For detection of inflammatory cytokine expression at the site of injection, mRNA expression in the tissue samples were examined as follow. RNAs were extracted using a Sepasol-RNA I Super G RNA-isolation kit (Nacalai Tesque). After the removal of DNA contamination by DNase I (Nippon Gene), the total RNA was reverse-transcribed with ReverTra Ace qPCR RT Master Mix (TOYOBO) to synthesize cDNA. Quantitative real-time PCR (qRT-PCR) was performed using THUNDERBIRD SYBR qPCR Mix (TOYOBO) and StepOnePlus (Thermo fisher scientific) for the presence of IL-1β/6/12α/17A, TNF-α, and MIP1/2.
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