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    • 5. 发明授权
    • Sequential step method for sequencing and identifying polynucleotides
    • 用于测序和鉴定多核苷酸的顺序步骤
    • US5908755A
    • 1999-06-01
    • US950709
    • 1997-10-15
    • Rajan KumarPaul Heaney
    • Rajan KumarPaul Heaney
    • C12N15/09C12Q1/68G06F17/30C12P19/34G01N33/48
    • C12Q1/6869Y10T436/143333
    • In one aspect, described are methods of sequencing of polynucleotides using a sequential step procedure. The methods of the invention begin with the provision of a single-stranded polynucleotide template that is annealed with a primer, forming a template-primer complex. The methods of the invention can provide for adding discrete nucleotides by a polymerase to the template-primer complex, where the added nucleotides can be labeled. The methods of the invention can involve the identification of a polynucleotide or polynucleotides having a contiguous non-redundant string or a superimposed non-redundant string pattern. The detection of the presence of a non-redundant contiguous string can be used, for example, to identify a particular gene. Alternatively, for example, if the non-redundant contiguous string is not unique to a particular gene, the string can be used to form a DNA library that can then be searched, for example, with a second string. Similarly, a superimposed non-redundant string pattern can be used, for example, to identify a gene or to search a DNA library. Such strings can be used, for example, in annealing reactions and in computer searches of a database having a catalog of sequenced polynucleotides.
    • 在一个方面,描述了使用顺序步骤程序测序多核苷酸的方法。 本发明的方法开始于提供与引物退火的单链多核苷酸模板,形成模板引物复合物。 本发明的方法可以提供通过聚合酶将离散的核苷酸加入到模板 - 引物复合物中,其中加入的核苷酸可被标记。 本发明的方法可以涉及鉴定具有连续非冗余字符串或叠加的非冗余字符串模式的多核苷酸或多核苷酸。 例如,可以使用非冗余连续字符串的存在的检测来鉴定特定基因。 或者,例如,如果非冗余连续字符串对于特定基因不是唯一的,则该字符串可以用于形成DNA文库,然后可以例如用第二个字符串来搜索。 类似地,可以使用叠加的非冗余字符串模式,例如识别基因或搜索DNA文库。 这样的串可用于例如退火反应和计算机搜索具有测序多核苷酸目录的数据库。