| 摘要 |
<p>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.</p> |
