Selective Isolation of DNA or RNA Using Single-Stranded DNA Affinity Latex Particles.
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.
Imai
T
Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama, 227, Japan
Sumi
Y
Hatakeyama
M
Fujimoto
K
Kawaguchi
H
Hayashida
N
Shiozaki
K
Terada
K
Yajima
H
Handa
H