Fig. 4

Performance of primer sets with different delta G cut-off values. A Genome alignment of a single primer, T13k_1. The nonspecific amplicon shows a strong binding stability on chr6. B, C, D, E, and F represent the amplification of the under − 13, − 12 ~ − 13, − 11 ~ − 12, − 10 ~ − 11, and over − 9 kcal/mol sets, respectively; S refers to the human genome DNA sample, and N refers to the water negative control. G Verification of primer sets with different delta G cut-off values