Method for determining DNA nucleotide sequence

A method for sequencing DNA comprising

• obtaining nucleic acid transcripts while DNA fragments comprising a target DNA fragment sequence are being amplified, by combining a DNA polymerase and a RNA polymerase with
• a DNA fragment (a-1) comprising the target DNA fragment sequence
• wherein the DNA fragment (a-1) comprises a sequence accepting formation of a nick and on at least one strand, a promoter sequence for a RNA polymerase,
• a primer (G1) comprising
• a primer sequence for one strand of the target DNA fragment and a restriction enzyme recognition sequence,
• a primer (G2) comprising
• a primer sequence for the other strand of the target DNA fragment and a sequence accepting formation of a nick,
• provided that at least one of the primers G1 and G2 comprises the promoter sequence for the RNA polymerase,
• deoxyribonucleoside-5′ triphosphates comprising
• dATP, dGTP, dCTP and dTTP or derivatives thereof,
• ribonucleoside-5′ triphosphates comprising
• ATP, GTP, CTP and UTP or derivatives thereof, and
• 3′ deoxyribonucleoside-5′ triphosphates comprising
• 3′ ATP, 3′ GTP, 3′ CTP and 3′ UTP or derivatives thereof,
• forming a nick at a site of the DNA fragment (a-1) accepting formation of a nick;
• separating the nucleic transcripts;
• and
• determining the nucleic acid sequence of said nucleic acid transcript.

A method of target DNA sequencing comprising

• hybridizing primers B1, B2, G1, and G2 to a DNA fragment (a-1) comprising a target DNA fragment, to produce a DNA fragment (a-2);
• obtaining nucleic acid transcripts, while simultaneously amplifying DNA fragments (A-2) comprising the target DNA fragment and performing an extension reaction, by combining
• a DNA polymerase and a RNA polymerase
• a DNA fragment (a-2)
• the primer B1,
• the primer G2,
• dNTP derivatives,
• NTP derivatives and
• at least one kind of 3′ dNTP derivatives, and
• forming a nick at a site of the DNA fragment (a-2) restriction enzyme recognition sequence;
• separating the nucleic acid transcripts and
• determining the nucleic acid sequence of said nucleic acid transcripts.