Author: Su, Xiaoping; Qian, Cheng; Zhang, Qian; Hou, Jin; Gu, Yan; Han, Yanmei; Chen, Yongjian; Jiang, Minghong; Cao, Xuetao
Title: miRNomes of haematopoietic stem cells and dendritic cells identify miR-30b as a regulator of Notch1 Document date: 2013_12_6
ID: 4vo7n6nh_38
Snippet: Luciferase assays. The Notch1 3 0 UTR luciferase reporter construct was prepared by amplifying the mouse Notch1 mRNA 3 0 UTR sequence by PCR and cloned into pMIR-Report construct (Ambion, Austin, TX, USA). Three sequences including S1 or S2 sites were individually or in combination cloned into pGL3 luciferase reporter plasmid (Promega). HEK293 cells described above were co-transfected with 80 ng luciferase reporter plasmid, 40 ng pRL-TK-Renilla-l.....
Document: Luciferase assays. The Notch1 3 0 UTR luciferase reporter construct was prepared by amplifying the mouse Notch1 mRNA 3 0 UTR sequence by PCR and cloned into pMIR-Report construct (Ambion, Austin, TX, USA). Three sequences including S1 or S2 sites were individually or in combination cloned into pGL3 luciferase reporter plasmid (Promega). HEK293 cells described above were co-transfected with 80 ng luciferase reporter plasmid, 40 ng pRL-TK-Renilla-luciferase plasmid, and indicated RNAs (final concentration: 20 nM). After 24 h, Luciferase activities were measured using Dual-Luciferase Reporter Assay System (Promega) according to the man' and in Dulbecco's modified Eagle's medium (DMEM ufacturer's instructions. Data was normalized for transfection efficiency by dividing Firefly luciferase activity with that of Renilla luciferase. All primers are listed in Supplementary Data 3.
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