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EGFP miSFIT Lineage Barcoding Library
(Pooled Library #229136)

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  • Purpose

    The EGFP miSFIT Lineage Barcoding library is a transcribed lineage barcoding vector library. It can be used to create a lentiviral barcoding pool that is robustly expressed in human PSC-derived hematopoietic cells. The library comprises a bi-directional promotor driving expression of two transcripts, a transmembrane reporter (tNGFR), and an EGFP mRNA that contains a 16-mer randomized barcode upstream of the poly(A) tail. It is possible to recover the barcode sequences and transcriptional information from standard 3’ single cell RNA capture reactions, including 10x genomics.

  • Vector Backbone

    EGFP-miSFIT-1x (Plasmid #124690)

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 229136 EGFP miSFIT Lineage Barcoding Library 1 $704 Add to Cart
Available to Academic and Nonprofits Only

Library Details

  • Unique barcodes
    Theoretically, there are 416 = 4,294,967,296 possible unique inserts, but the library contains at least 500,000 unique barcodes.
  • Size of barcodes
    24 nt
  • Lentiviral generation
    3rd

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
    50 ng/µL

Resource Information

Depositor Comments

The library comprises a bi-directional promotor (PGK/minCMV) driving expression of two transcripts, a transmembrane reporter (tNGFR) upstream of a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), and an EGFP mRNA that contains a 16-mer randomized barcode upstream of the poly A tail.

Figure 1: EGFP miSFIT Lineage Barcoding library construct.

Please visit https://doi.org/10.1101/2023.10.06.561277 (Link opens in a new window)for bioRxiv preprint.

How to cite this pooled library ( Back to top )

These pooled libraries 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:

    EGFP miSFIT Lineage Barcoding library was a gift from Peter Zandstra (Addgene #229136 ; http://n2t.net/addgene:229136 ; RRID:Addgene_229136)

  • For your References section:

    Tracking the gene expression programs and clonal relationships that underlie mast, myeloid, and T lineage specification from stem cells. Michaels YS, Major MC, Bonham-Carter B, Zhang J, Heydari T, Edgar JM, Siu MM, Greenstreet L, Vilarrasa-Blasi R, Kim S, Castle EL, Forrow A, Ibanez-Rios MI, Zimmerman C, Chung Y, Stach T, Werschler N, Knapp DJHF, Vento-Tormo R, Schiebinger G, Zandstra PW. Cell Syst. 2024 Dec 18;15(12):1245-1263.e10. doi: 10.1016/j.cels.2024.11.001. Epub 2024 Nov 29 PubMed 39615483
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