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WormBase Tree Display for Construct: WBCnstr00022774

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Name Class

WBCnstr00022774Summary[UNC-9::GFP]
GeneWBGene00006749
Fusion_reporterGFP
Construction_summaryTo generate a full-length unc-9::gfp construct, the 14.7-kb UNC-9A+B PCR product (representing the 5' end of unc-9) and the unc-9::gfp plasmid were digested separately with KpnI, digests electrophoresed through low-melt agarose (SeaPlaque GTG agarose, FMC, Rockland, ME, USA), and appropriate fragments purified by phenol extraction and ethanol precipitation. Approximately 0.6-1 ug of each fragment was used in a 20-ul ligation reaction, and 1ul of ligation mix used for long-range PCR amplification with primers UNC-9A + UNC-9D to produce a final 18.4-kb product. Reaction conditions followed the manufacturer's recommendations, with an annealing temperature of 63 deg C, an initial 15 minute extension time, and 10+18cycles.
Used_forTransgene_constructWBTransgene00022992
ReferenceWBPaper00033138