Embodiments of the present disclosure relate to methods of preparation of templates for polynucleotide sequencing. In particular, the disclosure relates to linearization of clustered polynucleotides in preparation for sequencing by cleavage of one or more first strands of double-stranded polynucleotides immobilized on a solid support by a transition metal complex, for example, a palladium complex or a nickel complex. Further disclosure relate to linearization of clustered polynucleotides by cleaving one or more second strands of double double-stranded polynucleotides immobilized on a solid support comprising azobenzene linker by Na
2
S
2
O
4
. Nucleotides and oligonucleotides comprising a 3′ phosphate moiety blocking group, and methods of removing the same using a fluoride reagent are also disclosed.
本公开的实施例涉及用于多核苷酸测序模板制备的方法。具体而言,本公开涉及通过过渡
金属络合物(例如
钯络合物或
镍络合物)裂解固定在固体支持物上的双链多核苷酸的一条或多条第一链,以便为测序做准备。进一步公开涉及通过Na2S2O4裂解固定在含有
偶氮苯联结剂的固体支持物上的双链多核苷酸的一条或多条第二链来线性化团簇化的多核苷酸。还公开了含有3'
磷酸酯基团阻断基团的核苷酸和寡核苷酸,以及使用
氟化试剂去除这些基团的方法。