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. 2023 Nov 14;145(47):25789–25796. doi: 10.1021/jacs.3c09497

Figure 2.

Figure 2

Selection conditions. (a) Parallelized single-round aptamer screening demands stringent binding conditions to partition functional and nonfunctional sequences. Cartoon representation of an evaluation of the binding and wash conditions for a starting library tested against S1. (b) Summary of the selection conditions evaluated to increase the separation of active and inactive sequences. Buffer 1: 25 mM Tris (pH 8.0) and 150 mM NaCl. Buffer 2: 10 mM HEPES (pH 7), 150 mM NaCl, 3 mM EDTA, 0.05% Tween 20, 0.5 mg/mL BSA, and 0.05 mg/mL ssDNA. Washing conditions refer to selection buffer supplemented with (a) 1 M NaCl, (b) 0.5 M urea, (c) 1 M urea, (d) 2 M urea, and (e) no additive. Library refolding involves heating to 65 °C and cooling on ice. T/L refers to target/library molar ratio and [L]/[E] denotes the molar ratio of the starting library and elution sequences.