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. 2016 Oct 1;27(5):197–208. doi: 10.1089/hgtb.2016.059

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© Huan-Ting Lin et al. 2016; Published by Mary Ann Liebert, Inc.

This Open Access article is distributed under the terms of the Creative Commons Attribution Noncommercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited.

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Figure 1. — Primer design and target specificity. (a) Bidirectional arrows indicate differences in primer design targeting either codon-optimized (CO) (white letters) or genomic (gray letters) CYBB sequences. (b) Amplification curves for the detection of CO CYBB and the puromycin resistance gene (PURO) (pink). As a control, primer probes targeting genomic CYBB (green) were included within the same reaction samples. The plasmid pAlpha.SIN.EFS.gp91s.F2A.PURO.T2A.ΔLNGFR.oPRE was diluted to 10 and 100 times. The horizontal pink line represents C_t (threshold cycle) values. NTC, no-template control; RFU, relative fluorescence units. (c) Tabular summary of CO CYBB and PURO detection alongside respective C_t values. N/A, not applicable (indicates no detection); NTC, no-template control.