Inclusion of self-inhibiting nucleotides in the hybridization arm improves contrast. (A) Experimental scheme used to measure fluorescence emergence. (B) Comparison of the fluorescence emergence for the Mango I Beacon and Mango II Beacon with a 4-nt inhibitory domain (MIB + 4i and MIIB + 4i, respectively) and without this domain (MIB and MIIB, respectively). Time courses were performed as follows: TO1-Biotin solution (D, 100 nM final, added at t = 5 min) is incubated with a Mango Beacon construct (B, 50 nM final concentration, added at t = 10 min) and then a trigger sequence added (T, 100 nM final concentration, added at t = 70 min). (C) Cartoon showing the design of the MIIB + ni construct, where n refers to the number of base pairs of hybridization, as indicated by the arrow.
