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. 2016 Nov 28;45(6):3487–3502. doi: 10.1093/nar/gkw1141

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© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 1. — The principal scheme of the Flowseq experiment. Presented are the steps of library construction, transformation, sorting and sequencing. (A) Cloning of the randomized DNA fragment into pRFPCER reporter vector in front of CER gene. RFP gene retains its constant 5΄-UTR. (B) Electroporation of entire plasmid library into E. coli cells. (C) Separation of cells on the basis of CER/RFP fluorescence by cell sorter. (D) Collection of cell pools (F1–F8) according to CER/RFP fluorescence ratio. (E) DNA extraction and amplification of randomized region followed by next generation sequencing.