The pGL3 control vector plasmid (Promega GmbH, Mannheim, Germany), encoding for the firefly luciferase, was transformed into chemical competent E.coli cells (One Shot Top 10, Life Technologies, Darmstadt, Germany) according to the manufacturer’s protocol. Bacterial colonies were amplified in ampicillin-containing medium, and the plasmid cDNA was purified using the QIAGEN Plasmid Plus Midi Kit (QIAGEN GmbH, Hilden, Germany) according to the manufacturer’s high yield protocol. The plasmid content was determined with a Nanodrop 2000™ spectrophotometer (Thermo Scientific, Wilmington, USA), and the plasmid cDNA stocks were stored at −20°C.
Pgl3 control vector plasmid
Manufactured by Promega
Sourced in Germany
The PGL3 control vector plasmid is a laboratory tool used in gene expression studies. It contains a reporter gene that can be used to measure the activity of a promoter or regulatory sequence. The plasmid is designed to provide a baseline for comparison in experiments involving the evaluation of gene expression.
Automatically generated - may contain errors
Lab products found in correlation
Nanodrop 2000, by Thermo Fisher Scientific (2 mentions)
Plasmid plus midi kit, by Qiagen (1 mentions)
One shot top10, by Thermo Fisher Scientific (1 mentions)
Dual glo luciferase assay, by Promega (1 mentions)
Dharmafect duo transfection reagent, by Thermo Fisher Scientific (1 mentions)
Glomax luminometer, by Promega (1 mentions)
2 protocols using pgl3 control vector plasmid
1
Plasmid Amplification and Purification
2
Screening Artificial miRNA Regulators
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Artificial miRNAs were screened for luciferase reporter repression activity in 293T cells in triplicate. Ten picomoles artificial miRNAs were reverse co-transfected into 5 × 103 cells along with 50 ng pLightSwitch_3UTR reporter plasmid (SwitchGear Genomics) and 25 ng pGL3-Control Vector plasmid (Promega, E1741) using 2.5 ul DharmaFECT Duo Transfection Reagent (Thermo Scientific) in 100 ul total volume. The 3′ UTR reporter plasmids used were PC (S803754), GLS (S812820), GAPDH (S801378) and the empty vector (S890005). Twenty-four hours post-transfection, cells were assayed for luciferase activity using the Dual-Glo Luciferase Assay (Promega) according to the manufacturer's protocol, except that 75 ul of each reagent was used. Firefly and Renilla luciferase activity were measured on a GloMax Luminometer (Promega) using a 1 s integration time with an 18 min incubation time following addition of each reagent. Analysis of luciferase reporter data was performed with R and the plyr, qvalue and ggplot2 packages (26 –29 ).
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