Wild-type (BTG2 and hsa_circ_0069382 genes are not mutated) and mutant vectors (BTG2 and hsa_circ_0069382 genes are mutated) were constructed with pmirGlo, (GenePharma, Shanghai, China), respectively. A positive control vector (pmirGlo, GenePharma, Shanghai, China) of the miR-15a-5p inhibitor and a negative control vector (pmirGlo, GenePharma, Shanghai, China) of NC-FAM were constructed. GP-transfect mate (GenePharma, Shanghai, China) was used as the transfection reagent. We co-transfected the miR-15a-5p mimics and reporter gene vectors into 293 T cells. A dual-luciferase reporter gene detection system kit (Promega, USA) and Synergy HTx multifunctional microplate reader (Biotek, USA) were used to obtain the data.
Dual luciferase reporter gene detection system kit
Manufactured by Promega
Sourced in United States
The Dual-luciferase reporter gene detection system kit is a tool used to quantify the activity of two different luciferase reporter genes simultaneously. It provides a rapid and sensitive method for the measurement of firefly and Renilla luciferase activities in transfected cells or cell extracts.
Automatically generated - may contain errors
2 protocols using dual luciferase reporter gene detection system kit
1
Dual-Luciferase Assay for miR-15a-5p
2
Dual-Luciferase Assay for miRNA-lncRNA Interaction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
This experiment used the Dual-Luciferase reporter gene detection system kit (Promega Corporation), which was used according to the manufacturer's instructions. GP-transfect-Mate transfection reagent (Shanghai GenePharma Co., Ltd.) was used for transfection. Cells were seeded in 12-well plates at a density of 5×105 cells per well for 24 h before transfection. Reporter plasmids [pGP-miRGLO-Firefly luciferase-Renilla luciferase, containing lncKRT16P6, the GATAD2A 3′ untranslated region (UTR), or the corresponding mutant (MUT) sequences] were designed by Shanghai GenePharma Co., Ltd. 293(T) cells were cotransfected with a reporter plasmid and either miR-3180 mimic or the NC mimic (Shanghai GenePharma Co., Ltd.). After 48 h, luciferase activity was measured using a dual-luciferase reporter assay system (Promega Corporation) according to the manufacturer's protocol. Firefly luciferase activity was normalized to Renilla luciferase activity.
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!