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    • 3. 发明申请
    • NUCLEIC ACID SEQUENCING BY ADAPTATOR LIGATION
    • 核酸序列由适配器定位
    • WO1998015652A1
    • 1998-04-16
    • PCT/GB1997002734
    • 1997-10-06
    • BRAX GENOMICS LIMITEDSCHMIDT, GunterTHOMPSON, Andrew, Hugin
    • BRAX GENOMICS LIMITED
    • C12Q01/68
    • C12Q1/6872C12Q2565/518C12Q2525/191C12Q2521/313
    • A method for sequencing nucleic acid, which comprises: (a) obtaining a target nucleic acid population comprising nucleic acid fragments in which each fragment is present in a unique amount and bears at one end a sticky end sequence of predetermined length and unknown sequence, (b) protecting the other end of each fragment, and (c) sequencing each of the fragments by (i) contacting the fragments with an array of adaptor oligonucleotides under hybridisation conditions, each adaptor oligonucleotide bearing a label, a sequencing enzyme recognition site, and a known unique base sequence of same predetermined length as the sticky end sequence, the array containing all possible base sequences of that predetermined length; removing any unhybridised adaptor oligonucleotide and recording the quantity of any hybridised adaptor oligonucleotide by detection of the label, then repeating the cycle, until all of the adaptors in the array have been tested; (ii) contacting the hybridised adaptor oligonucleotides with a sequencing enzyme which binds to the recognition site and cuts the fragment to expose a new sticky end sequence which is contiguous with or overlaps the previous sticky end sequence; (iii) repeating steps (i) and (ii) for a sufficient number of times and determining the sequence of the fragment by comparing the quantities recorded for each sticky end sequence.
    • 一种用于测序核酸的方法,其包括:(a)获得包含核酸片段的靶核酸群,其中每个片段以唯一的量存在,并且在一端具有预定长度和未知序列的粘性末端序列( b)保护每个片段的另一端,和(c)通过以下步骤对每个片段进行测序:(i)在杂交条件下将片段与衔接子寡核苷酸阵列接触,每个具有标记的衔接子寡核苷酸,测序酶识别位点和 与粘性末端序列相同的预定长度的已知唯一碱基序列,该阵列包含该预定长度的所有可能碱基序列; 去除任何未经杂交的接头寡核苷酸并通过检测标记物记录任何杂交的接头寡核苷酸的量,然后重复该循环,直到阵列中的所有适配器已被测试; (ii)使杂交的接头寡核苷酸与测序酶接触,所述测序酶结合识别位点并切割片段以暴露与先前粘性末端序列连续或重叠的新粘性末端序列; (iii)重复步骤(i)和(ii)足够的次数,并且通过比较每个粘性末端序列记录的量来确定片段的序列。