RNA fluorescence in situ hybridization using 3-cyanovinylcarbazole modified oligodeoxyribonucleotides as photo-cross-linkable probes.
Photo-cross-linkable fluorescent oligodeoxyribonucleotides having 3-cyanovinylcarbazole nucleoside were applied to fluorescence in situ hybridization (FISH) based 16S rRNA detection in Escherichia coli cells. As the photo-cross-linked probe/rRNA hybrid was stable under the denaturing condition, decrease of the fluorescence signal through the washing process was prevented. The thermally irreversible hybridization property also enabled stable hybridization with the structured region on the target RNA, and facilitated design of the sequence for the FISH probe. Further development of the method might contribute to quantitative and stable FISH staining.
School of Materials Science, Japan Advanced Institute of Science and Technology, 1-1 Asahi-dai, Nomi, Ishikawa 923-1292, Japan.
Kenzo
K
Fujimoto
School of Materials Science, Japan Advanced Institute of Science and Technology, 1-1 Asahi-dai, Nomi, Ishikawa 923-1292, Japan.
Kei
K
Toyosato
School of Materials Science, Japan Advanced Institute of Science and Technology, 1-1 Asahi-dai, Nomi, Ishikawa 923-1292, Japan.
Shigetaka
S
Nakamura
School of Materials Science, Japan Advanced Institute of Science and Technology, 1-1 Asahi-dai, Nomi, Ishikawa 923-1292, Japan.
Takashi
T
Sakamoto
D002227:Carbazoles / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D015335:Molecular Probes; D009838:Oligodeoxyribonucleotides / Q000737:chemistry*; D055668:Photochemical Processes; D012313:RNA / Q000737:chemistry*
D002227:Carbazoles; D015335:Molecular Probes; D009838:Oligodeoxyribonucleotides; D012313:RNA
16S rRNA; 3-Cyanovinylcarbazole nucleoside; Fluorescence in situ hybridization; Photo-cross-link