Takara Bio Inc.

Takara Shuzo Co., Inc.

Johns Hopkins University

Rutgers, the State University of New Jersey

Hiroyuki Mukai

Takashi Uemori

Ikunoshin Kato

Kiyozo Asada

Yoshimi Sato

Kazue Miyake

Junko Yamamoto

Kiyoyuki Matsumura

Tomoko Fujita

Osamu Takeda

Nariaki Takatsu

Kazue Nishiwaki

Eiji Kobayashi

Shigekazu Hokazono

Fumitsugu Hino

Nana Shimada

Hiroaki Sagawa

Michio Hagiya

Keiko Kubo

Masayori Inouye

Tatsuji Enoki

Hikaru Takakura

Jun Tomono

Naoyuki Awazu

Hideyo Yamaguchi

Masanari Kitagawa

Toshiharu Ohba

Yoshie Yoshikawa

Christian B Anfinsen

Sangita Phadtare

Toshihiro Shodai

Kenneth Laderman

Satoshi Yanagi

Mariko Moriyama

Yuko Nakabayashi

Haruhisa Sawaragi

Mariko Okawa

Ling Zhu

Kanako Usui

Tomoo Inden

Yuki Ueda

Tooru Suzuki

A composition for use in amplifying cDNA synthesized by a reverse transcription reaction and detecting RNA that serves as a template of the reverse transcription reaction, the composition containing a thermostable DNA polymerase, a thermostable ribonuclease H, and an intercalating dye. Since the composition of the present invention can suppress the influences to the nucleic acid amplification reaction by RNA that serves as a template for cDNA synthesis, the composition is useful in the detection of RNA, and more useful in quantification of RNA having a desired sequence by real-time RT-PCR.

Composition for detection of RNA

Provided are a mismatch-specific cleavage reaction using a novel heat-resistant mismatch nuclease, a method for removing errors in a nucleic acid amplification reaction using the mismatch nuclease, a method for inhibiting the amplification of a nucleic acid having a specific base sequence during a nucleic acid amplification reaction, and a method for detecting a nucleic acid having a single-base polymorphic mutation using this inhibition method.

Method for using heat-resistant mismatch endonuclease

Provided are the following: a method, for improving reactivity of an acid synthesis reaction, comprising a step for adding an &#x3c9;-amino acid to a reaction solution; a composition, for a nucleic acid synthesis reaction, comprising DNA polymerase, reaction buffer, at least one primer, at least one deoxyribonucleoside triphosphate, and an &#x3c9;-amino acid; and a reaction buffer, for a nucleic acid synthesis reaction, comprising an &#x3c9;-amino acid.

Method for improving nucleic acid synthesis reaction

Provided are the following: a method, for improving reactivity of a nucleic acid synthesis reaction, comprising a step for adding an &#x3c9;-amino acid to a reaction solution; a composition, for a nucleic acid synthesis reaction, comprising DNA polymerase, reaction buffer, at least one primer, at least one deoxyribonucleotide triphosphate, and an &#x3c9;-amino acid; and a reaction buffer, for a nucleic acid synthesis reaction, comprising an &#x3c9;-amino acid.

The present invention pertains to a method for modifying a nucleic acid contained in a sample comprising a step for bringing the sample into contact with a nucleic acid-modifying agent in the presence of an acidic polysaccharide and/or a nucleotide; and a method for selectively detecting a nucleic acid derived from living cells contained in the sample, comprising the following steps: (a) a step for modifying a nucleic acid contained in a sample according to the method for modifying a nucleic acid contained in a sample, which includes a step for bringing the sample into contact with a nucleic acid-modifying agent in the presence of an acidic polysaccharide and/or a nucleotide; and (b) a step for selectively detecting an unmodified nucleic acid from the sample after step (a). The present invention further pertains to a kit and composition for use in these methods.

Method for modifying nucleic acids

The present invention pertains to: a method for modifying a nucleic acid contained in a sample, the method including a step for bringing the sample into contact with a nucleic acid-modifying agent in the presence of an acidic polysaccharide and/or a nucleotide; and a method for selectively detecting a nucleic acid derived from living cells contained in the sample, the method including the following steps: (a) a step for modifying a nucleic acid contained in a sample according to the method for modifying a nucleic acid contained in a sample, which includes a step for bringing the sample into contact with a nucleic acid-modifying agent in the presence of an acidic polysaccharide and/or a nucleotide; and (b) a step for selectively detecting an unmodified nucleic acid from the sample after step (a). The present invention further pertains to a kit and composition for use in these methods.

The present invention provides cold shock protein-containing compositions for improved DNA synthesis reactions with improved reactivity, methods for synthesizing DNA using such compositions, kits for use in such methods, and DNA compositions yielded by such methods. The present invention further provides cold shock protein-containing compositions for the identification of endoribonuclease cleavage sites, methods for identifying endoribonuclease cleavage sites using such compositions, and kits for use in such methods.

Cold shock protein compositions and methods and kits for the use thereof

An object of the present invention is to provide a simple and useful method for producing an RNA-containing probe for detecting a target nucleotide, a simple and useful method, device, and system for processing nucleotide sequence information, and a simple and useful method for detecting a target nucleotide. The present invention provides a method for processing nucleotide sequence information, the method comprising the step of generating partial nucleotide sequences which has 7 to 14 nucleotides and a Tm value of 25 to 40&#xb0; C. and in which a target nucleotide or a nucleotide adjacent to the target nucleotide is located at a position between 3 and 5 nucleotides from the 3' or 5&#x2032; end. The method according to the present invention is useful for simply and efficiently producing an RNA-containing probe for detecting a target nucleic acid, without the basis of researchers' experiences or guess, and are extremely useful not only in the field of genetic engineering, but also in the field of medical research.

Method for producing RNA-containing probe for detecting a target nucleotide

A method for accurately and easily detecting a synthetic siRNA, for example, a siRNA in which the 3' end is DNA, and a kit used for the method are provided. The present invention relates to a method for detecting a siRNA in which the 3&#x2032; end is DNA, comprising: (a) adding polydeoxyadenosine to the 3&#x2032; DNA end of at least one strand of the siRNA to be detected to produce a polydeoxyadenosine-added RNA; (b) annealing a polydeoxythymidine primer having a tag sequence at its 5&#x2032; side to the polydeoxyadenosine-added RNA and synthesizing DNA from the primer by a reverse transcription; and (c) detecting the DNA synthesized in (b).

Synthetic siRNA detection method

A method for synthesizing cDNA characterized by preparing a reaction solution that does not allow an endodeoxyribonuclease to show its activity, without thermal deactivation of the endodeoxyribonuclease or removal of the endodeoxyribonuclease, and carrying out a reverse transcription reaction, wherein the reaction solution contains a treated sample and a reverse transcriptase, the treated sample being formed by treating a sample comprising RNA and DNA with the endodeoxyribonuclease to degrade DNA in the sample. The method and the kit for synthesizing cDNA of the present invention are widely useful in genetic engineering fields.

Method for synthesizing cDNA

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 polypeptide having a high fidelity DNA polymerase activity and thus being useful as a genetic engineering reagent; a gene encoding this polypeptide; a method of producing the polypeptide; and a method of amplifying a nucleic acid by using the polypeptide.

Polypeptides having DNA polymerase activity

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Otsu, Japan

Hiroyuki Mukai