Rabies Barcode Pooled Libraries
(Pooled Libraries #227671, #227672)

• Purpose:

  The Rabies Barcode libraries are a collection of glycoprotein (G)-deleted rabies virus (RVdG) genome plasmids that carry an error-correctable barcode compatible with 3’ capture single cell or single nuclei RNA sequencing readout (snRNA-seq).

  The pSADB19-dTomato barcode library (#227671) is used for the production of SADB19 strain G-deleted rabies virus.

  The pCVSN2c-MCP-oScarlet-KASH-6xMS2 barcode library (#227672) is used for the production of CVS N2c strain G-deleted rabies virus. Barcodes in this library are compatible with snRNA-seq.

• Vector Backbone:

  • Library #227671: pSADdeltaG-BFP (Plasmid #32639)
  • Library #227672: RabV CVS-N2c(deltaG)-mCherry (Plasmid #73464)

• Depositing Labs: Tomasz Nowakowski, Cathryn Cadwell
• Publication: Shin and Urbanek et al. Biorxiv. 2024 Dec 11. doi: 10.1101/2024.10.01.616167.

Library Details

• Number of barcodes: Theoretical diversity of 125,000,000
• Virus type/Generation: Rabies Virus
• Insert size:
  
  • Library #227671: 910 bp
  • Library #227672: 2087 bp

Library Shipping

Each library is delivered in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze/thaws.

• Volume: ∼10 µL
• Concentration: 250 ng/µL

Resource Information

• Protocols:
  • Library Amplification Protocol (DOCX, 19 KB)
  • Library Sequencing Protocol (DOCX, 254 KB)
• Depositor Data:
  • List of Primer Sequences (XLSX, 9 KB)
  • Library Representation (GEO)
• Scripts:
  • Deconvolution algorithms for analyzing NGS data (GitHub)

Terms and Licenses

Academic/Nonprofit Terms:

• UBMTA
• Evrogen Limited Use Label License for FPs
• Takara Bio Limited Use Label License (formerly Clontech)

Industry Terms:

• Not Available to Industry

Trademarks:

• Zeocin® is an InvivoGen trademark.

Depositor Comments

We recommend ethanol precipitation of library samples and re-elution in water before electroporation, as salts present in the sample buffer (10 mM Tris-Cl, pH 8.5) can reduce electroporation efficiency. Please, find more information in the library amplification protocol provided.

Please visit https://doi.org/10.1101/2024.10.01.616167 for bioRxiv preprint.

Figure 1: Location of barcode cloning site in the rabies genome for the Rabies pSADB19-dTomato Barcode library (left) and the Rabies pCVSN2c-MCP-oScarlet-KASH-6xMS2 Barcode library (right). Creative Commons Attribution License .