File:MIT2012 In vitro RNA SD with diagram small.png

Strand displacement in vitro. The input strand (Part:BBa_K779118) can attach via the toehold T to the reporter complex (Part:BBa_K779117:Part:BBa_K779116) and go through toehold-mediated strand displacement as described by [http://www.sciencemag.org/content/332/6034/1196.abstract Qian et al. 2011]. This results in a dsROX complex (see above) and a free quencher strand (Part:BBa_K779117). The dsROX complex is fluorescent, whereas the original reporter is not. Negative control is reporter, but with a scramled-sequence strand (Part:BBa_K779119) as an input. After 2 hours, a roughly 10-fold increase in fluorescence is observed. Measurements carried out on a Tecan Safire II plate reader (see [http://2012.igem.org/Team:MIT/MaterialsAndMethods#iv2bio protocol]).