Nucleic acid amplification method producing uracil-containing amplification products

A nucleic acid amplification method, comprising the steps of

• performing a first nucleic acid amplification using a target nucleic acid contained in a sample as a template, and
• performing a second nucleic acid amplification with forward and reverse primers, using the amplification product produced in the first nucleic acid amplification step as a template,
• wherein
• in the first nucleic acid amplification, a DNA polymerase that selectively amplifies a nucleic acid not comprising a uracil base is used to produce an amplification product that does not comprise a uracil base, the DNA polymerase being derived from an archaebacteria, and
• in the second nucleic acid amplification, (i) dUTP and/or a primer comprising a uracil base, and (ii) a thermostable DNA polymerase derived from Thermus aquaticus and capable of amplifying a nucleic acid comprising a uracil base are used to produce an amplification product, wherein said amplification product comprises double-stranded nucleic acid having uracil on both strands thereof.

A nucleic acid amplification method, comprising the steps of

• performing a first nucleic acid amplification using a target nucleic acid contained in a sample as a template,
• performing a second nucleic acid amplification with forward and reverse primers, using the amplification product produced in the first nucleic acid amplification step as a template, and
• detecting an amplification product of the second nucleic acid amplification step using an intercalator or a labeled probe that binds to the amplification product of the second nucleic acid amplification,
• wherein
• in the first nucleic acid amplification, a DNA polymerase that selectively amplifies a nucleic acid not comprising a uracil base is used to produce an amplification product that does not comprise a uracil base, the DNA polymerase being derived from an archaebacteria, and
• in the second nucleic acid amplification, (i) dUTP and/or a primer comprising a uracil base, and (ii) a thermostable DNA polymerase derived from Thermus aquaticus and capable of amplifying a nucleic acid comprising a uracil base are used to produce an amplification product, wherein said amplification product comprises double-stranded nucleic acid having uracil on both strands thereof.