URL: http://www.addgene.org/124296
Proper Citation: RRID:Addgene_124296
Insert Name: His6 tag - TEV protease site - T7 tag - Streptavidin (15-159) - superfolder GFP
Organism: Synthetic
Bacterial Resistance: Kanamycin
Vector Backbone Description: Backbone Marker:Novagen; Backbone Size:5237; Vector Backbone:pET29b; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: T7 promoter so expression requires DE3 strain. Pilot experiments using E. coli BL21(DE3) indicate 18C expression overnight gives higher yields than 37C (for 4 hours). The fusion protein was soluble and purified by Ni-NTA chromatography using 50 mM Tris pH 8.0, 300 mM NaCl, 10/300 mM imidazole. Size exclusion chromatography indicated that the fusion protein was tetrameric in Tris-buffered saline; no aggregates were present.
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Source: Addgene (RRID:SCR_002037)
Description: Non-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. Facilitates archiving and distributing DNA-based research reagents and associated data to scientists worldwide. Repository contains over 65,000 plasmids, including special collections on CRISPR, fluorescent proteins, and ready-to-use viral preparations. There is no cost for scientists to deposit plasmids, which saves time and money associated with shipping plasmids themselves. All plasmids are fully sequenced for validation and sequencing data is openly available. We handle the appropriate Material Transfer Agreements (MTA) with institutions, facilitating open exchange and offering intellectual property and liability protection for depositing scientists. Furthermore, we curate free educational resources for the scientific community including a blog, eBooks, video protocols, and detailed molecular biology resources.
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