Figure 2.

CycLiC Sequencing. (A) Top: Image of microarray spot (strongest signal) for first base addition and corresponding histogram showing the intensity of the four labeled degenerate ON libraries. Bottom: Image of microarray spot and histogram showing intensities for all four degenerate ON libraries, after cleavage and addition of the second base. NB data is normalized to strongest signal. Second base addition data is not corrected for template loss or for residual signal from cycle 1. (B) Polyacrylamide gel analysis of iterative cycles of ligation and cleavage carried out in solution. The first lane shows the product of first base ligation and cleavage. As the cycles of ligation and cleavage are repeated the size of the fragment increases base by base. The first three ligations were carried out with ONs specific to the sequence in the template while the fourth ligation was carried out with the OL corresponding to the complementary base in the template. The sketch below the gel illustrates the primer growing in length by one base at a time.