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. 2019 Jun 21;2(3):172–185. doi: 10.1089/crispr.2019.0009

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© Nicholas C. Huston, et al. 2019; Published by Mary Ann Liebert, Inc.

This Open Access article is distributed under the terms of the Creative Commons License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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FIG. 1. — Barcoded library of targets (BLT) can measure Cas9 specificity at scale in vitro. (A) Schematic of barcoded library of targets experiment. Cleavage first by Cas9 and then by EcoRV allows measurement of cleavage efficiency by next-generation sequencing. Targets are identified by association with a unique molecular identifier (UMI). (B) Theoretical maximum library size and empirical distributions of mismatched targets analyzed in the data. (C) Cleavage efficiency measurements in BLT are highly reproducible across experiments.