Engineering a Robust DNA Circuit for the Direct Detection of Biomolecular Interactions by Ang Yan Shan

Author:Ang Yan Shan
Language: eng
Format: epub, pdf
ISBN: 9789811321887
Publisher: Springer Singapore

The shorter hairpin stem was more effective in reconfiguring itself in response to α-thrombin binding and so, we retained the 5 nt length while adding a “clamp” domain in Design iii and iv to improve hairpin stability without promoting the “unlocking” process. We also considered placing the aptamer sequence at the end of the hairpin stem in Design v (Lane 16 and 17) to avoid the kinetically slow process of hairpin opening which turned out to be an ineffective strategy.

The only design that remained locked in absence of α-thrombin yet triggered HCR in presence of α-thrombin was Design iii (Lane 12 and 13). The “clamp” domain was the crucial element in buffering the transient exposure of the “key” domain when the ends of the stem “breathed” as in Design i and Design ii. The clamp length greatly influenced the outcome of this strategy as excessive length led to overly stable hairpin as observed for Design iv (Lanes 14 and 15). This hindered the hairpin opening process such that no HCR occurred in presence of α-thrombin. Thus, we selected Design iii for the rest of this study which gave a reasonable signal-to-background ratio of approximately 3.5 (Fig. 4.1b).

Download

Copyright Disclaimer:
This site does not store any files on its server. We only index and link to content provided by other sites. Please contact the content providers to delete copyright contents if any and email us, we'll remove relevant links or contents immediately.