Takara Bio Inc.

Takara Shuzo Co., Inc.

Osamu Takeda

Ikunoshin Kato

Kiyozo Asada

Hiroyuki Mukai

Takashi Uemori

Yoshimi Sato

Kazue Miyake

Tomoko Fujita

Hiroaki Sagawa

Michio Hagiya

Eiji Kobayashi

Tatsuji Enoki

Jun Tomono

Junko Yamamoto

Hideyo Yamaguchi

Mariko Okawa

Mariko Moriyama

Haruhisa Sawaragi

A DNA synthesis reaction-enhancer comprising at least one kind selected from the group consisting of acidic substances and cationic complexes; a DNA synthesis method in which during a DNA synthesis reaction a reaction is carried out in the presence of the above enhancer by using DNA polymerase; a DNA synthesis reaction composition comprising the above enhancer; a DNA synthesis reaction composition comprising two or more kinds of DNA polymerases each having 3'&#x2192;5&#x2032; exonuclease activity; a DNA synthesis method in which during a DNA synthesis reaction two or more kinds of DNA polymerases each having 3&#x2032;&#x2192;5&#x2032; exonuclease activity are used; a kit for use in in vitro DNA synthesis, comprising two or more kinds of DNA polymerases each having 3&#x2032;&#x2192;5&#x2032; exonuclease activity; and a kit for use in in vitro DNA synthesis, wherein the kit comprises the DNA synthesis reaction-enhancer and DNA polymerase. According to the present invention, DNA synthesis can be carried out at an efficiency more excellent as compared to conventional DNA synthesis reaction.

Method for synthesizing DNA

A convenient and effective method for amplifying a nucleic acid sequence characterized by effecting a DNA synthesis reaction in the presence of chimeric oligonucleotide primers; a method for supplying a large amount of DNA amplification fragments; an effective method for amplifying a nucleic acid sequence by combining the above method with another nucleic acid sequence amplification method; a method for detecting a nucleic acid sequence for detecting or quantitating a microorganism such as a virus, a bacterium, a fungus or a yeast; and a method for detecting a DNA amplification fragment obtained by the above method in situ.

Method for amplifying nucleic acid sequence

A DNA synthesis method with a shortened time period required for DNA synthesis by polymerase chain reaction (PCR), characterized in that a DNA polymerase is used in an amount effective for providing more than 10 ng of amplified DNA fragments of about 2 kb per 50 &mgr;l of a reaction mixture, when PCR is carried out under the following conditions (A) and (B): (A) reaction mixture: 50 &mgr;l volume of a reaction mixture comprising DNA polymerase, 1 ng of genomic DNA from and 10 pmol each of primers Eco-1 and Eco-2 (nucleotide sequences of the primers Eco-1 and Eco-2 being shown in SEQ ID NOs: 10 and 11 of Sequence Listing, respectively); and having a composition suitable for the DNA polymerase; and (B) reaction conditions: 35 cycles of PCR, wherein one cycle consists of 99&deg; C., 1 second-66&deg; C., 7 seconds; a kit for DNA synthesis usable for the DNA synthesis method; and an article of manufacture of a PCR agent. According to the present invention, the procedures in the genetic engineering studies and industries involved with PCR can be speeded up.

This invention is directed to a gene sequence for secretory aspartic proteinase (SEQ ID No. 1) which is specific to pathogenic Candida yeast, to a highly sensitive method for detection of said gene in a sample solution and a kit used therefor.

Gene encoding aspartic acid secretory proteinase of Candida albicans and

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Hikone, Japan

Osamu Takeda