Total RNA was extracted from 3D4/21 cells using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) and the first-strand cDNA was synthesized using a First-Strand Synthesis System (Transgen, Beijing, China) according to the manufacturer’s instructions. The relative gene expression was analyzed by real-time RT-qPCR carried out on qTOWER3G Real-time PCR system (Analytik Jena AG, Jena, Germany). Relative gene expression levels were calculated using the comparative cycle threshold (CT) method according to the manufacturer’s protocol (Applied Biosystem, Foster City, CA, USA). All data presented are relatively quantitative, normalized to the β-actin and analyzed using GraphPad Prism 8.0 software. All primers used for real-time RT-qPCR are listed in Supplementary Table S2.
First strand synthesis system
Manufactured by Transgene
Sourced in China
The First-Strand Synthesis System is a laboratory product designed for the synthesis of complementary DNA (cDNA) from total RNA or mRNA. It provides the necessary reagents and protocols for performing the first-strand cDNA synthesis step, which is a crucial part of many molecular biology techniques, such as RT-PCR, cDNA library construction, and gene expression analysis.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using first strand synthesis system
1
Gene Expression Analysis by RT-qPCR
2
Quantitative Gene Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Quantification of the relative levels of gene expression was performed using quantitative reverse transcription-PCR (qRT-PCR). Total RNA from 3D4/21 cells was extracted using TRIzol reagent (TaKaRa), and the first-strand cDNA was synthesized using a first-strand synthesis system (TransGen, China) according to the manufacturer’s instructions. The relative levels of gene expression were analyzed by using the qTower3 G in vitro diagnostic (IVD) fluorescence quantitative PCR system (Jena, Germany) using TransStart top green qPCR super mix (TransGene) with three-step amplification. The data were calculated based on the cycle threshold (2−ΔΔCT) method, and the data were normalized to the expression level of the β-actin gene. All of the primer pairs used for quantitative real-time PCR are listed in Table 3 .
3
Quantitative Real-Time PCR Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
First-strand cDNA was synthesized from purified RNAs of 3D4/21 cells or HEK293T cells using a First-Strand Synthesis System (Transgen, Beijing, China) according to the manufacturer's instructions. The relative gene expression was analyzed by qRT-PCR that was performed on an ABI 7500 Real-time PCR system (Applied Biosystems, Foster City, CA, USA). The comparative cycle threshold (CT) method was used to calculate the relative gene expression levels according to manufacturer's protocol (Applied Biosystem). All data presented was relatively quantitative, based on the mRNA level of the endogenous gene β-actin and analyzed using GraphPad Prism 6.0 software. All of the primers pairs used for quantitative real-time PCR are listed in Table 2 .
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!