cDNA was generated from 750 ng of total RNA using MultiScribe Reverse Transcriptase (Thermo Fisher Scientific, 4311235) and oligo(dT) primer (Thermo Fisher Scientific, N8080128). Quantitative PCR was performed using the CXF96 Touch Real-Time PCR Detection System (BioRad, 1855195) with iQ SYBR Green Supermix (BioRad, 1708880). An annealing temperature of 60°C was used for each primer set. Samples were run in triplicate and normalized to GAPDH. The ΔΔCt method was used to calculate relative gene expression between samples [85 (link)]. Primer sequences for KRT14, KRT5, FLG, SFN, and GAPDH are listed in S10 Table .
Cxf96 touch real time pcr detection system
Manufactured by Bio-Rad
Sourced in United States
The CXF96 Touch Real-Time PCR Detection System is a compact, easy-to-use instrument designed for real-time PCR applications. It features a touchscreen interface and a 96-well block format for efficient sample processing. The system is capable of detecting multiple fluorescent dyes and supports various real-time PCR workflows.
Automatically generated - may contain errors
2 protocols using cxf96 touch real time pcr detection system
1
cDNA Synthesis and qPCR Analysis of Skin Markers
2
Quantifying Wound Healing Gene Expression
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Real-time polymerase chain reaction (qPCR) was performed to determine the changes in gene expression. The genes were selected corresponding to the cytokines involved at different phases of wound healing. Amplification was performed using qPCR (CXF96 Touch Real-Time PCR Detection System, Bio-Rad, United States) under the following conditions; denaturation at 95°C for 1 min, annealing at 58°C for 1 min (40 cycles) and extension at 72°C for 1 min. GAPDH was taken as an internal control. The genes analyzed are presented in Table 1 .
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