This invention relates to the field of nucleic acid chemistry, more specifically to the field of compositions of matter that comprise triphosphates of modified 2′-deoxynucleosides and oligonucleotides that are formed when these are appended to the 3′-end of a primer, wherein said modifications comprise NH
2
moiety attached to their 3′-hydroxyl group and a fluorescent species in a form of a tag affixed to the nucleobase via a linker that can be cleaved. Such compositions and their associated processes enable and improve the sequencing of oligonucleotides using a strategy of cyclic reversible termination, as outlined in U.S. Pat. No. 6,664,079. Most specifically, the invention concerns compositions of matter that are 5′-triphosphates of ribo- and 2′-deoxyribonucleosides carrying detectable tags and oligonucleotides that might be derived from them. The invention also concerns processes wherein a DNA polymerase, RNA polymerase, or reverse transcriptase synthesizes said oligonucleotides via addition of said triphosphates to a primer.
本发明涉及核酸
化学领域,更具体地涉及包含经过修饰的2'-去氧核苷酸
三磷酸盐和寡核苷酸的物质组成,当它们附加到引物的3'-端时形成,其中所述修饰包括连接到它们的3'-羟基上的NH2基团和通过可断链的连接器固定在核碱基上的荧光物种。这些组成物及其相关的过程使得并改进了使用循环可逆终止策略对寡核苷酸进行测序,如美国专利号6,664,079所概述的。最具体地,本发明涉及载有可检测标记的
核糖核苷酸和2'-去氧
核糖核苷酸的5'-
三磷酸盐以及可能从它们中派生出的寡核苷酸的物质组成。本发明还涉及
DNA聚合酶、RNA聚合酶或
反转录酶通过向引物添加所述
三磷酸盐来合成所述寡核苷酸的过程。