As previously described (24 (link)), total RNA was isolated from mouse skin or lung tissue using Trizol (Invitrogen Life Technologies) according to the manufacturer's instructions. cDNA was prepared using the Reverse Transcription System (Promega). The expression of related genes was measured using gene-specific primers (shown in Table S1 ) with SYBR Green (SYBR Premix Ex Taq RT-PCR kit, Takara) and the 7500 real-time PCR system analyzer (Applied Biosystems). GAPDH expression was used as the endogenous control to normalize the sample data. Relative expression levels were calculated using the 2−ΔΔCt method.
7500 real time pcr system analyzer
Manufactured by Thermo Fisher Scientific
The 7500 real-time PCR system analyzer is a laboratory instrument designed for quantitative real-time PCR analysis. It is capable of detecting and measuring specific DNA sequences in real-time during the amplification process.
Automatically generated - may contain errors
2 protocols using 7500 real time pcr system analyzer
1
RNA Isolation and Gene Expression Analysis
2
PBMC Isolation and Gene Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Peripheral blood samples were obtained from patients and controls as described previously [23 (link)]. We isolated PBMCs in Ficoll-Paque Plus (GE Healthcare) by density gradient centrifugation. We used Trizol (Invitrogen Life Technologies) to isolate RNA from PBMCs and a Reverse Transcription System (Promega) to obtain cDNA. The relative expression of genes was measured by gene-specific primers (shown in Additional file 2 : Table S2) with SYBR Green (SYBR Premix Ex Taq RT-PCR kit; Takara) and the 7500 real-time PCR system analyzer (Applied Biosystems).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!