Method and composition for identifying cognate nucleotides in a Sequencing By Binding™ procedure, wherein one or more labeled nucleotides are detected in ternary complexes but never incorporated. Labeled nucleotides can be incorporable nucleotides that contact preformed blocked primed template nucleic acids. Alternatively, labeled nucleotides are labeled non-incorporable nucleotides. Labeled nucleotides, including labeled non-incorporable nucleotides, can be detected in ternary complexes in the same reaction mixture that incorporates a reversible terminator nucleotide to create a blocked primed template nucleic acid. Detection of ternary complexes can take place in the presence of a catalytic metal ion.
在 Sequencing By Binding™ 程序中识别同源核苷酸的方法和组合物,其中在三元复合物中检测到一种或多种标记核苷酸,但从未与之结合。标记的核苷酸可以是可结合的核苷酸,与预先形成的阻断引物模板核酸接触。或者,标记的核苷酸是标记的不可结合核苷酸。标记的核苷酸,包括标记的不可结合核苷酸,可以在同一反应混合物中的三元复合物中检测到,该反应混合物结合了可逆的终止子核苷酸,生成了阻断引物的模板核酸。三元复合物的检测可在催化
金属离子存在的情况下进行。