pPB_mU6_enh-gRNA_Puro-T2A-BFP
(Plasmid
#235571)
-
PurposeEnhanced guide RNA expression & genomic integration. Target gRNA sequences are cloned in via the BstXI and BlpI sites.
-
Depositing Lab
-
Depositing OrganizationEuropean Molecular Biology Laboratory (EMBL), Rome
Located in Italy -
Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 235571 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 235571-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 235571-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
-
Vector backbonepPB-U6
-
Vector typeCRISPR
Growth in Bacteria
-
Bacterial Resistance(s)Ampicillin, 100 μg/mL
-
Growth Temperature30°C
-
Growth Strain(s)NEB Stable
-
Copy numberHigh Copy
Gene/Insert
-
Gene/Insert namesgRNA backbone
-
gRNA/shRNA sequenceUnspecified
-
SpeciesSynthetic
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site Unknown (unknown if destroyed)
Resource Information
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
Drives expression of a sub-cloned single guide RNA (gRNA) with enhanced scaffold to target epigenetic editing to a desired locus. gRNA are cloned using the BlpI and BstXI sites. This is a piggyBAC plasmid that integrates into the genome and can be selected for via Puromycin resistance. An additional tagBFP insert is expressed constituitively and indirectly indicates presence of the gRNA.
DNA (Catalog # 235571-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
-
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.
DNA (Catalog # 235571-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.
-
For your Materials & Methods section:
pPB_mU6_enh-gRNA_Puro-T2A-BFP was a gift from Jamie Hackett (Addgene plasmid # 235571 ; http://n2t.net/addgene:235571 ; RRID:Addgene_235571) -
For your References section:
Systematic epigenome editing captures the context-dependent instructive function of chromatin modifications. Policarpi C, Munafo M, Tsagkris S, Carlini V, Hackett JA. Nat Genet. 2024 Jun;56(6):1168-1180. doi: 10.1038/s41588-024-01706-w. Epub 2024 May 9. 10.1038/s41588-024-01706-w PubMed 38724747
