Toyo Boseki Kabushiki Kaisha

Hiroaki Inoue

Yoshihisa Kawamura

Masao Kitabayashi

Tadayuki Imanaka

Bunsei Kawakami

Masahiro Takagi

Masaaki Morikawa

Taku Arakawa

Yutaka Takarada

Shuji Shibata

In a method for amplifying the specific nucleic acid sequence, a highly specific amplification which has low possibility of non-specific hybridization can be carried out. Highly stable reagents, the activities of which do not decrease in case of supply and storage, are also provided. Thermostable enzymes are used as RNA dependent DNA polymerase, DNA dependent DNA polymerase, DNA dependent RNA polymerase and ribonuclease H, which are required for the amplification system based on replicated RNA. Especially, it is preferable that a thermostable enzyme derived from,which has RNA dependent DNA polymerase activity, DNA dependent DNA polymerase activity and ribonuclease H activity, and thermostable DNA dependent RNA polymerase are used together. By this method, inactivation of enzymes are prevented by using thermostable enzymes, and the amplification can be carried out without sequential addition of enzymes.

Method for amplifying and detecting of target nucleic acid sequence using thermostable enzyme

A nucleic acid amplifying enzyme having a short reaction time and high fidelity is provided. The enzyme of this invention is a thermostable DNA polymerase having a nucleic acid extension rate of at least 30 bases per second and a 3&prime;-5&prime; exonuclease activity. Also provided are a method and kit for amplifying nucleic acid.

Kit for amplifying nucleic acid

A nucleic acid amplifying enzyme having a short reaction time and high fidelity is provided. The enzyme of this invention is a thermostable DNA polymerase having a nucleic acid extension rate of at least 30 bases per second and a 3'-5' exonuclease activity. Also provided are a method and kit for amplifying nucleic acid.

Methods of amplification using a thermostable DNA polymerase from the

Thermostable DNA polymerase from a hyperthermophilic archaeon strain KOD1

In a method for amplifying the specific nucleic acid sequence, a highly specific amplification which has low possibility of non-specific hybridization can be carried out. Highly stable reagents, the activities of which do not decrease in case of supply and storage, are also provided. Thermostable enzymes are used as RNA dependent DNA polymerase, DNA dependent DNA polymerase, DNA dependent RNA polymerase and ribonuclease H, which are required for the amplification system based on replicated RNA. Especially, it is preferable that a thermostable enzyme derived from Thermus thermophilus which has RNA dependent DNA polymerase activity, DNA dependent DNA polymerase activity and ribonuclease H activity, and thermostable DNA dependent RNA polymerase are used together. By this method, inactivation of enzymes are prevented by using thermostable enzymes, and the amplification can be carried out without sequential addition of enzymes.

Method for amplifying and detecting of target nucleic acid sequence

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Tsuruga, Japan

Hiroaki Inoue