公开(公告)号：EP1182267A4

公开(公告)日：2004-09-15

申请号：EP01917739

申请日：2001-03-30

申请人： TOYOTA MOTOR CO LTD ,  GENESIS RES INST INC

发明人： KONDOW TAMOTSU ,  MAFUNE FUMITAKA ,  TAKEDA YOSHIHIRO

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6869 ,  C12Q2565/518 ,  C12Q2563/107

摘要： A method of determining the base sequence of a nucleic acid by detecting a single dye molecule, which involves the following steps: (1) fixing to a solid surface a nucleic acid molecule or a primer having a sequence complementary to a part of the sequence thereof; (2) annealing the above nucleic acid molecule or primer with the corresponding primer or nucleic acid molecule; (3) supplying a solution containing DNA polymerase and a dye-labeled dNTP or RNA polymerase and a dye-labeled NTP to the fixed nucleic acid molecule and reacting the nucleotide with the 3'-end of the primer to thereby allowing the polymerase to incorporate the nucleotide forming a base pair with the base opposite to the reaction site; (4) detecting the dye-labeled dNTP or NTP thus bonded; (5) disrupting the dye molecule of the bonded dye-labeled dNTP or NTP; (6) repeating the procedures of the steps (3) to (5) while successively changing the type of the dye-labeled dNTP or NTP so as to bond the dNTP or NTP forming a base pair with the nucleotide of the nucleic acid molecule; and (7) determining the base sequence of the nucleic acid molecule based on the dNTP or NTP thus bonded.