A kit and methods for nucleic acid sequencing of single molecules by polymerase synthesis

(57) Summary book invention relates the nucleic acid in single molecule arrangement to how to have been improved sequence determination and for carrying out genotype determination. This method includes detection of the single molecule of the fluorescent label PPi portion emitted from NTP, when a polymerase extension product is produced. When all four nucleotides are added simultaneously, addition of a gradual nucleotide is unnecessary. Arrangement information is generated succeeding the case where polymerase takes in all four nucleotides continuously into the nucleic acid NA chain currently elongated. Since a single molecule is separately observed in the present invention, there is no loss of synchronization. Analysis of a single molecule enables again use of NA fragmentation taken out directly from the living thing.