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URL: http://www.addgene.org/226799
Proper Citation: RRID:Addgene_226799
Insert Name: nanos2
Organism: Danio rerio
Bacterial Resistance: Kanamycin
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:2640; Vector Backbone:pDONR-P2R-P3; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: Cloned by Maya Pahima. The putative zebrafish nos2 3'utr was cloned from zebrafish ovary gDNA using primers (3’-ATCAACCAATGCTGAATCTTTTTATAAAAAATTGAA-5' and 3’-AAATAAAATATATCAGGAGGAAACAAATATATCAGG-5') and inserted into the pCR4 cloning vector using the TOPO™ TA Cloning Kit (Thermo Fisher, K457502) for stabilization and sequencing (pCR4-nos2,3'utr). To insert the nos2,3’utr sequence into a Gateway Cloning vector, the region was amplified with attB primers (3'-GGGGACAGCTTTCTTGTACAAAGTGGTATATCAACCAATGCTGAATCTTTTTATAAAAAATTGAA-5' and 3'-GGGGACAACTTTGTATAATAAAGTTGTAAATAAAATATATCAGGAGGAAACAAATATATCAGG-5') and a BP reaction was performed with the Gateway BP Clonase II Enzyme mix (Invitrogen, 11789020) to recombine the amplicon into pDONR-P2R-P3 (Invitrogen, 12537-023) and produce p3E-nos2,3’utr.
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Source: Addgene
