Fig. 3 Analysis of FACS-based screening efficiency by competitive allele-specific TaqMan PCR. Analysis of FACS-based screening efficiency by competitive.

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Fig. 3 Analysis of FACS-based screening efficiency by competitive allele-specific TaqMan PCR. Analysis of FACS-based screening efficiency by competitive allele-specific TaqMan PCR. (A) The RBS-ATG spacing technique was used to create two populations of cells: (i) normal FutA(+) cells and (ii) FutA(+) cells with a weakened FutA activity resulting from reduced FutA expression [FutA(+)-RBS cells]. Cell mixtures of FutA(+) and FutA(+)-RBS were prepared and applied to one round of FACS sorting. The unsorted and sorted variant pools were quantified using competitive allele-specific TaqMan PCR, and then, enrichment factors were calculated according to FutA(+) cell ratios before and after sorting. (B) Flow cytometric screening of FutA(+) and FutA(+)-RBS cells. (C) Percentage of FutA(+) cells increased after sorting. Yumeng Tan et al. Sci Adv 2019;5:eaaw8451 Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).