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PurposeGFP-based reporter construct pMHG-W57* capable of detecting dose-dependent drug-induced PTC readthrough both by fluorescence microscopy and flow cytometry.
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Depositing Lab
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Depositing OrganizationVanderbilt University
Located in United States of America -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 72850 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 72850-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 72850-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepMONOhygro-gfp (pMHG)
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Backbone manufacturerInvivogen
- Backbone size w/o insert (bp) 4728
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Modifications to backboneSite-directed mutagenesis of GFP tryptophan 57 codon to a stop codon (W57⁄) (G171A, mRNA). An opal stop codon (TGA) was introduced using the following primers (forward, G CTG CCA GTG CCT TGA CCA ACT CTG GTG AC; reverse, GTC ACC AGA GTT GGT CAA AGG CAC TGG CAG C).
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Vector typeMammalian Expression
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Selectable markersHygromycin
Growth in Bacteria
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Bacterial Resistance(s)Hygromycin, 200 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberUnknown
Gene/Insert
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Gene/Insert nameGFP
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SpeciesH. sapiens (human)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
DNA (Catalog # 72850-D50)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 50 μg
- Concentration 1 μg/μL
- Pricing $495 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
DNA (Catalog # 72850-D100)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 100 μg
- Concentration 1 μg/μL
- Pricing $585 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
pMHG-W57* reporter was a gift from Daniel Liebler (Addgene plasmid # 72850 ; http://n2t.net/addgene:72850 ; RRID:Addgene_72850) -
For your References section:
A Reporter System for Translational Readthrough of Stop Codons in Human Cells. Halvey PJ, Liebler DC, Slebos RJ. FEBS Open Bio. 2012 Dec;2:56-59. Epub 2012 Apr 17. 10.1016/j.fob.2012.04.004 PubMed 22563532
