精確序列資訊及修飾鹼基位置確認的方法

工研院創新前瞻技術研究計畫

潘詔智,范振業,邱創汎,簡虹琪,陳惠玲

本發明揭露確認核酸樣本的序列及/或核酸樣本中修飾鹼基的位置之方法，該核酸樣本以具有已知序列的核酸插入物之環狀分子呈現，該方法包含獲取至少兩個插入樣本單元的序列資訊。在一些實施例中，本發明方法包括使用環狀雙股連結分子獲取序列資訊。在一些實施例中，本發明方法包括藉由比較核酸樣本序列與核酸插入物之已知序列，計算核酸插入物序列的分數，及根據緊鄰於核酸樣本序列的重複序列上游或下游之一個或兩個插入物的序列分數，接受或拒絕該核酸樣本序列的重複序列。

Disclosed herein are methods of determining the sequence and/or positions of modified bases in a nucleic acid sample present in a circular molecule with a nucleic acid insert of known sequence comprising obtaining sequence data of at least two insert-sample units.  In some embodiments, the methods comprise obtaining sequence data using circular pair-locked molecules.  In some embodiments, the methods comprise calculating scores of sequences of the nucleic acid inserts by comparing the sequences to the known sequence of the nucleic acid insert, and accepting or rejecting repeats of the sequence of the nucleic acid sample according to the scores of one or both of the sequences of the inserts immediately upstream or downstream of the repeats of the sequence of the nucleic acid sample.