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 | |
| Publication status | Published - 1996 Jan 15 |
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Keywords
- affinity latex particles
- complementary RNA
- hybridization selection
- single-stranded DNA
ASJC Scopus subject areas
- Colloid and Surface Chemistry
- Physical and Theoretical Chemistry
- Surfaces and Interfaces
Cite this
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
}
TY - JOUR
T1 - Selective isolation of DNA or RNA using single-stranded DNA affinity latex particles
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
PY - 1996/1/15
Y1 - 1996/1/15
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
UR - http://www.scopus.com/inward/record.url?scp=0029951923&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0029951923&partnerID=8YFLogxK
U2 - 10.1006/jcis.1996.0027
DO - 10.1006/jcis.1996.0027
M3 - Article
AN - SCOPUS:0029951923
VL - 177
SP - 245
EP - 249
JO - Journal of Colloid and Interface Science
JF - Journal of Colloid and Interface Science
SN - 0021-9797
IS - 1
ER -