Figure 2.

Nick-labeling scheme. (A) Recognition sequence of nicking endonuclease Nb.BbvCI and the labeling scheme. After nicking (arrow), a fluorescent nucleotide terminator is incorporated at the nicking site (green T) as the native T is displaced. (B) The specificity of the nick-labeling scheme was demonstrated with synthetic oligos containing the Nb.BbvC I recognition sequence. In eight parallel experiments containing all possible combinations of synthetic oligos with or without the Nb.BbvC I site and the four dye-terminators, only the synthetic oligo with the Nb.BbvC I site was nick-labeled with the T-terminator, as monitored by a steep rise in fluorescence polarization value.