The mRNA expression of CHI3L1, COX-2, GPX-1 and CAT in the ankle joint tissue was measured using qRT-PCR as described previously (Huang et al. 2019 (link); Izu et al. 2019 (link)). Total RNA was extracted from frozen ankle joint with Trizol Reagent (Takara, Dalian, China) following the manufacturer’s instructions. cDNA synthesis was performed with Frist Strand cDNA Synthesis Kit (Takara, Dalian, China). SYBR Green qPCR Master Mix kit (Thermo Scientific, Wilmington, DE) was used to quantify the mRNA levels. The qPCR was performed in triplicates, the parameter of RT-PCR amplification reaction as follows: 40 cycles of 95 °C for 10 s (denaturation) and 72 °C for 30 s (annealing/extension) using the primer sequences (Table 1 ). The levels of mRNA expression were normalized to endogenous control gene GAPDH.
Frist strand cdna synthesis kit
Manufactured by Takara Bio
Sourced in China
The Frist Strand cDNA Synthesis Kit is a laboratory product designed to perform the first-strand cDNA synthesis step in reverse transcription reactions. It provides the necessary components to convert RNA into complementary DNA (cDNA) for further downstream applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using frist strand cdna synthesis kit
1
Quantitative Analysis of Ankle Joint Gene Expression
2
Cytokine Expression in Kidney
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mRNA expression of TNF-α, IL-1β, IL-6 and MCP-1 in kidney tissues was explored utilizing RT-qPCR. Trizol Reagent (Takara, Dalian, China) was utilized to harvest total RNA, followed by cDNA synthesis by Frist Strand cDNA Synthesis Kit (Takara, Dalian, China), as guided by the instructions. Thereafter, relative mRNA expression was quantified with the help of SYBR Green qPCR Master Mix kit (Thermo Scientific, Wilmington, DE) in triplicates. The parameters of RT-qPCR amplification reaction were as follows: 40 cycles of 95°C for just 10 s (denaturation) and 72°C for about 30 s (annealing/extension). Other mRNA expression levels were normalized to that of GAPDH.
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!