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. 2016 Sep 2;6:32644. doi: 10.1038/srep32644

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Copyright © 2016, The Author(s)

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(a) Optimization of heat-treatment temperature for screening. E. coli TOP10 (pET28a-P_tac) was a negative control while E. coli TOP10 (pET28a-P_tac-6pgdh) was a positive control. Colonies were treated at 23, 60, 70 and 80 °C for 1 h, respectively. The color changes of the colonies overlaid by the second agarose layer were examined. (b) Optimization of color development time and NAD⁺ concentration. Heat-treated colonies of E. coli TOP10 (pET28a-P_tac-6pgdh) was overlaid by the second layer containing 0, 0.1, 0.3 or 1 mM NAD⁺. The color change profiles of colonies were photographed at 0, 0.5, 1 and 2 h.