Selective isolation of DNA or RNA using single-stranded DNA affinity latex particles

Takeshi Imai; Yoshiteru Sumi; Mamoru Hatakeyama; Keiji Fujimoto; Haruma Kawaguchi; Nobuaki Hayashida; Kazuo Shiozaki; Kunihiko Terada; Hiroaki Yajima; Hiroshi Handa

doi:10.1006/jcis.1996.0027

Selective isolation of DNA or RNA using single-stranded DNA affinity latex particles

Takeshi Imai, Yoshiteru Sumi, Mamoru Hatakeyama, Keiji Fujimoto, Haruma Kawaguchi, Nobuaki Hayashida, Kazuo Shiozaki, Kunihiko Terada, Hiroaki Yajima, Hiroshi Handa

Department of Applied Chemistry

Research output: Contribution to journal › Article › peer-review

29 Citations (Scopus)

Abstract

We developed latex particles with a diameter of about 0.22 μm on which single-stranded (ss) DNA was covalently coupled to select or enrich its complementary DNA or mRNA. DNA was first covalently coupled to the latex particles in the double-stranded (ds) form with both blunt and protruding ends. More than 80% of the dsDNA was coupled through the ssDNA stretch at its protruding end. The presence of NaCl in the immobilization reaction severely inhibited DNA from coupling to the particles. The particles were then treated with alkali or heated to denature the dsDNA and sedimented by a brief centrifugation to yield ssDNA immobilized particles. They allowed the selective and efficient isolation of a desired RNA from total cellular RNA.

Original language	English
Pages (from-to)	245-249
Number of pages	5
Journal	Journal of Colloid And Interface Science
Volume	177
Issue number	1
DOIs	https://doi.org/10.1006/jcis.1996.0027
Publication status	Published - 1996 Jan 15

Keywords

affinity latex particles
complementary RNA
hybridization selection
single-stranded DNA

ASJC Scopus subject areas

Electronic, Optical and Magnetic Materials
Biomaterials
Surfaces, Coatings and Films
Colloid and Surface Chemistry

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10.1006/jcis.1996.0027

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Selective isolation of DNA or RNA using single-stranded DNA affinity latex particles. / Imai, Takeshi; Sumi, Yoshiteru; Hatakeyama, Mamoru; Fujimoto, Keiji; Kawaguchi, Haruma; Hayashida, Nobuaki; Shiozaki, Kazuo; Terada, Kunihiko; Yajima, Hiroaki; Handa, Hiroshi.

In: Journal of Colloid And Interface Science, Vol. 177, No. 1, 15.01.1996, p. 245-249.

Research output: Contribution to journal › Article › peer-review

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abstract = "We developed latex particles with a diameter of about 0.22 μm on which single-stranded (ss) DNA was covalently coupled to select or enrich its complementary DNA or mRNA. DNA was first covalently coupled to the latex particles in the double-stranded (ds) form with both blunt and protruding ends. More than 80% of the dsDNA was coupled through the ssDNA stretch at its protruding end. The presence of NaCl in the immobilization reaction severely inhibited DNA from coupling to the particles. The particles were then treated with alkali or heated to denature the dsDNA and sedimented by a brief centrifugation to yield ssDNA immobilized particles. They allowed the selective and efficient isolation of a desired RNA from total cellular RNA.",

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AU - Imai, Takeshi

AU - Sumi, Yoshiteru

AU - Hatakeyama, Mamoru

AU - Fujimoto, Keiji

AU - Kawaguchi, Haruma

AU - Hayashida, Nobuaki

AU - Shiozaki, Kazuo

AU - Terada, Kunihiko

AU - Yajima, Hiroaki

AU - Handa, Hiroshi

N1 - Funding Information: This work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Japan.

PY - 1996/1/15

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N2 - We developed latex particles with a diameter of about 0.22 μm on which single-stranded (ss) DNA was covalently coupled to select or enrich its complementary DNA or mRNA. DNA was first covalently coupled to the latex particles in the double-stranded (ds) form with both blunt and protruding ends. More than 80% of the dsDNA was coupled through the ssDNA stretch at its protruding end. The presence of NaCl in the immobilization reaction severely inhibited DNA from coupling to the particles. The particles were then treated with alkali or heated to denature the dsDNA and sedimented by a brief centrifugation to yield ssDNA immobilized particles. They allowed the selective and efficient isolation of a desired RNA from total cellular RNA.

AB - We developed latex particles with a diameter of about 0.22 μm on which single-stranded (ss) DNA was covalently coupled to select or enrich its complementary DNA or mRNA. DNA was first covalently coupled to the latex particles in the double-stranded (ds) form with both blunt and protruding ends. More than 80% of the dsDNA was coupled through the ssDNA stretch at its protruding end. The presence of NaCl in the immobilization reaction severely inhibited DNA from coupling to the particles. The particles were then treated with alkali or heated to denature the dsDNA and sedimented by a brief centrifugation to yield ssDNA immobilized particles. They allowed the selective and efficient isolation of a desired RNA from total cellular RNA.

KW - affinity latex particles

KW - complementary RNA

KW - hybridization selection

KW - single-stranded DNA

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Selective isolation of DNA or RNA using single-stranded DNA affinity latex particles

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Keywords

ASJC Scopus subject areas

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