Watermelon Library
(Pooled Library #155257)

• Purpose:

  The Watermelon library is a high complexity lentiviral barcode library for simultaneous tracing of clonal lineages as well as the transcriptional and proliferative state of each cell in the population. Lineage tracing is achieved by mapping a clone-specific expressed DNA barcode in the 3′ untranslated region of an mNeonGreen protein, and proliferation history is monitored by the dilution of a doxycycline (dox)-inducible H2B-mCherry transgene.

• Vector Backbone:

  Watermelon backbone

• Depositing Labs: Aviv Regev, Joan Brugge
• Publication: Oren et al Nature. 2021 Aug 11. pii: 10.1038/s41586-021-03796-6. doi: 10.1038/s41586-021-03796-6.

Library Details

• Number of Barcodes: 5x10⁶
• 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: ∼20µL
• Concentration: 50ng/uL

Resource Information

• Protocols:
  • Watermelon Library Generation Protocol (21.4 KB)
  • Electroporation Protocol (15.8 KB)
  • Dial Out PCR Protocol for Barcodes and miSeq Sequencing (22.3 KB)
  • Dial Out PCR for 10X WTA (19 KB)
• Depositor Data:
  • Watermelon Library Complexity (40.5 KB)
  • Sequence Logo of Barcode Area (102 KB)
  • Watermelon Library Sequence (Primers Marked) (26 KB)
  • 10X Dial Out Region (6 KB)
• Scripts:
  • Barcode Diversity Confirmation Script (3.1 KB)
• Terms and Licenses:
  • UBMTA
  • mNeonGreen Label License
  • Takara Bio Limited Use Label License (formerly Clontech)
  • Tet Systems Limited Use Label License

Depositor Comments

• The Watermelon system for simultaneous tracing of clonal lineages and transcriptional and proliferative state of individual cells.

The data and code related to this library is available on GitHub.

The file "Watermelon Library Sequence (Primers marked)" linked above provides an example of the plasmid sequence. Please see the Nature publication section "Cloning of the Watermelon library" to see how the final sequence compares to the Watermelon backbone (Plasmid #155258).

NOTE: Lentiviral vectors can recombine during library amplification, resulting in a ~6 kb and ~3.5 kb band. The recombined backbone product should not adversely affect library function as it does not contain lentiviral packaging sequences. If these additional bands represent a large percentage of the product after amplification, we recommend starting with a larger prep (gigaprep) from the original sample or gel purifying the amplified library to remove the recombinant band and performing another transformation with the purified sample.