...containing one or more mismatches. However, some ligases can ligate mismatches to a significant degree... Large purine:purine mismatches and most smaller pyrimidine:pyrimidine mismatches are typically worse ... pyrimidine:purine mismatches. Helix stability also plays some role, and mismatches with more hydrogen...of annealed mismatched substrates (expected to have a slightly lower Tm around the mismatch) over annealed...including substrates containing gaps or DNA base pair mismatches (8-11). For some applications, there cannot be...Ligase, can ligate nicks containing one or more mismatches near the ligation junction with high efficiency...ligated than those with few. For many ligases, G:T mismatches, with two hydrogen bonds and a base-pair size...