Moffat Lab - Barcode Libraries
(Pooled Libraries #179851, #179852)

• Purpose:

  This lentiviral barcoding library consists of 10⁶ unique barcodes. Each barcode contains a 12 bp degenerate region followed by two static bases (either C or G) and a unique 4 bp library code.

  The library was constructed by cloning the barcodes into a third generation lentiviral vector containing both a Puromycin selection cassette and the ZsGreen fluorescent marker.

  Cellular barcoding can be used to trace clonal dynamics of individual cells using stable, genetically distinct sequences inherited by all progeny.

• Vector Backbone:

  pLJM1

• Depositing Labs: Jason Moffat
• Publication: Rehman et al. Cell. 2021 Jan 7;184(1):226-242.e21. doi: 10.1016/j.cell.2020.11.018.

Library Details

• Number of barcodes: >10⁶
• Lentiviral Generation: 3rd

Library Shipping

This library is delivered as suspended DNA in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze-thaws.

• Volume: ∼40 µL
• Concentration: 50 ng/µL

Resource Information

• Protocols:
  • Library Amplification Protocol (17 KB)
  • Library Sequencing Protocol (134 KB)
  • Deconvolution Algorithm (15 KB)
• Scripts:
  • Barcode Deconvolution Script (3.5 KB)
• Depositor Data:
  • Barcode Index Primers (15 KB)
  • Library Complexity (238 KB)
• Terms and Licenses:
  • UBMTA
  • Institut Pasteur Label License for cPPT

Depositor Comments

Both are high diversity libraries identifiable by a unique 4 bp library code: BCL0 = ACGT; BLC1= CCAA. Either library can be used for an individual experiment.

The libraries have also been used and published in Chang et al. Cancer Discov. 2021 Dec 15. doi: 10.1158/2159-8290.CD-20-1265 (preprint available on bioRxiv: doi.org/10.1101/2020.08.28.273029).