Plasmid (100ng) and 50nmol/L miR-29a, miR-29b, miR-29c, or miR-100 (Dharmacon) were co-transfected using Lipofectamine 2000 (Invitrogen) in Plat-ES cells. Three replicates of each condition were used each time. Following 48 hr transfection, Renilla and luciferase activity were measured using a the Dual Glo Stop & Glo kit (Promega) and a FlexStation 3 (Molecular Devices) plate reader according to the dual luciferase reporting system instructions. Relative Renilla enzymatic activity was compared using the ratio of Renilla reniformis and firefly luciferase activity.
Mir 29a
Manufactured by Horizon Discovery
MiR-29a is a microRNA (miRNA) product offered by Horizon Discovery. miRNAs are small, non-coding RNA molecules that play a key role in gene expression regulation. The core function of MiR-29a is to act as a research tool for the study of miRNA-related biological processes.
Automatically generated - may contain errors
2 protocols using mir 29a
1
Plasmid and miRNA Dual Luciferase Assay
2
Transfection and Analysis of miR-29a in Cancer Cells
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Exponentially growing cancer cells were seeded in 6 well plates at 1×105 cells per well or 12 well plates at 5×104 per well and allowed to adhere overnight and transfected with indicated concentrations (10μM, 20μM) of control (GE Dharmacon, CN-001000-01) or miR-29a (GE Dharmacon, C-300504-07) mimics, or 1μM siRNA using siCTRL (GE Dharmacon, D-001810-10-05), siTFEB (GE Dharmacon, L-009798-00-0005), and siATG9A (GE Dharmacon, L-014294-01-0005) using DharmaFECT®1 (GE Dharmacon, T-2001-01) as per the manufacturer's protocol. Total protein or RNA was isolated at 24 hours post-transfection for western blot or qPCR analysis respectively as described above.
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