Perturb-Seq Guide Barcodes (GBC) Library
(Pooled Library #85968)

• Purpose:

  Barcode library for cloning gRNA sublibraries for single cell CRISPR analysis.

• Vector Backbone: pBA439 (Plasmid #85967)

• Depositing Labs: Jonathan Weissman
• Publication: Adamson et al Cell. 2016 Dec 15;167(7):1867-1882.e21. doi: 10.1016/j.cell.2016.11.048.

Library Details

• Lentiviral Generation: 3rd

Library Shipping

Each library is delivered in microcentrifuge tubes on blue ice. A tube's contents will not necessarily be frozen. For best results, minimize freeze/thaw cycles.

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

Resource Information

• Protocols:
  • Liquid Culture Re-Amplification Protocol (PDF, 70 KB)

Terms and Licenses

Academic/Nonprofit Terms:

• UBMTA
• Evrogen Limited Use Label License for FPs

Industry Terms:

• Not Available to Industry

Trademarks:

• Zeocin® is an InvivoGen trademark.

Depositor Comments

The Perturb-Seq Guide Barcodes (GBC) Library (also referred to as pBA571) is intended for cloning of individual protospacer sequences into the sgRNA expression cassette followed by isolation and verification of single barcoded expression vectors, as in the paper. It has not been optimized for pooled cloning. Of note, restriction digests indicate the presence of spurious DNA introduced during library construction from pBA439 that has not been observed to carry over into isolated sgRNA expression derivatives.

For amplification of the library, the use of high efficiency, electrocompetent cells is strongly advised. To ensure the maintenance of library diversity, calculate your transformation efficiency. 30x coverage of the library is recommended.

This library has been verified using individually cloned guide RNAs. Guide RNA protospacer sequences were individually cloned into the BstXI and BlpI sites by direct ligation. Protospacers and corresponding GBCs were then verified by sequencing. Final Perturb-seq vectors containing barcodes that introduce the conserved polyadenylation signal AATAAA should be discarded.