Figure 1

Structure and principle of bar-coded split tag. A. Schematic structure of bar-coded-split tag. This tag was a 16 bp oligo nucleotide consisting of a 13 bp basal region and a 3 bp bar-code region. A mismatch was introduced at the 8th nucleotide from the 5' end for the combination of a similar bar-code sequence to disturb annealing to the complementary sequence and to enhance selectivity. B. Principle of selective labeling with BStag. A complementary strand of PCR product amplified with an oligonucleotide primer harboring a 16 bp-long BStag sequence at its 5' end (center) anneals to the fluorescently labeled BStag primer with same sequence (upper panel). A fluorescently labeled BStag primer with similar sequence should not anneal to the complementary strand due to breakage of the annealing by the mismatched nucleotide at the bar-code sequence and the basal region (lower panel).