The entire 3’-UTR of SGK1 was amplified from human genomic DNA using PCR with the primer sequences: forward, 5’-ATAGCTAGCACCCTGTTAGGGCTTG and reverse, 5’-ATATCTCGAGCAACGGCTCTGACTG . The 3’-UTR was inserted into pmiRGLO Luciferase miRNA Target Expression Vector (Promega) and the sequence was confirmed by Sanger sequencing. Luciferase assays were conducted as previously reported following transfection with miRNA mimics or controls15 (link). β–Galactadase plasmid was co-transfected for normalization.
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Cloning and Luciferase Assay of SGK1 3'-UTR
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Corresponding Organization : Thomas Jefferson University
Other organizations : University of Alabama at Birmingham, Vanderbilt University
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Variable analysis
independent variables
- MiRNA mimics or controls
- Luciferase activity
- β–Galactadase plasmid for normalization
- Positive control: Not specified
- Negative control: miRNA mimics or controls
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