URL: http://www.addgene.org/63091
Proper Citation: RRID:Addgene_63091
Insert Name: expanded CGG repeats (99 repeats) within the 5'UTR of FMR1
Organism: Homo sapiens
Bacterial Resistance: Ampicillin
Vector Backbone Description: Vector Backbone:unknown; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Comments: Please note that Addgene NGS confirmed 4 fewer repeats (95x instead of 99x) for our stock of the plasmid, including one AGG and one TGG repeat. The depositor has noted that these differences should not affect the function of the polyglycine-containing protein. Should be amplified in STBL3 bacteria and grown at Room temperature to keep the repeats stable. This construct allows translation of CGG repeats into 99 glycine due to a near cognate ATG codon upstream of the repeat in the 5'UTR of FMR1 (an ACG codon that initiates weakly translation). These CGG are in frame with the EGFP tag. This plasmid should be verified via restriction analysis. Multiple colonies (at least 6) may need to be screened to isolate the correct construct. The plasmid should be screened for size every time it is amplified. We suggest a digest with HindIII and XhoI and running a 2% agarose gel. If correct expect 2 bands: 4218bp and 470bp. Smaller band is the insert.
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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.
URL: http://www.addgene.org