A quick and simple FISH protocol with hybridization-sensitive fluorescent linear oligodeoxynucleotide probes

Dan Ohtan Wang, Hitomi Matsuno, Shuji Ikeda, Akiko Nakamura, Hiroyuki Yanagisawa, Yasunori Hayashi, Akimitsu Okamoto

Research output: Contribution to journalArticlepeer-review

Abstract

Fluorescence in situ hybridization (FISH) is a powerful tool used in karyotyping, cytogenotyping, cancer diagnosis, species specification, and gene-expression analysis. Although widely used, conventional FISH protocols are cumbersome and time consuming. We have now developed a FISH method using exciton-controlled hybridization-sensitive fluorescent oligodeoxynucleotide (ECHO) probes. ECHO-FISH uses a 25-min protocol from fixation to mounting that includes no stringency washing steps. We use ECHO-FISH to detect both specific DNA and RNA sequences with multicolor probes. ECHO-FISH is highly reproducible, stringent, and compatible with other fluorescent cellular labeling techniques. The resolution allows detection of intranuclear speckles of poly(A) RNA in HeLa cells and dissociated hippocampal primary cultures, and mRNAs in the distal dendrites of hippocampal neurons. We also demonstrate detection of telomeric and centromeric DNA on metaphase mouse chromosomes. The simplicity of the ECHO-FISH method will likely accelerate cytogenetic and gene-expression analysis with high resolution.

Original languageEnglish (US)
Pages (from-to)166-175
Number of pages10
JournalRNA
Volume18
Issue number1
DOIs
StatePublished - Jan 2012

Keywords

  • Excitonic interaction
  • FISH
  • Probe
  • RNA
  • Thiazole orange

ASJC Scopus subject areas

  • Molecular Biology

Fingerprint

Dive into the research topics of 'A quick and simple FISH protocol with hybridization-sensitive fluorescent linear oligodeoxynucleotide probes'. Together they form a unique fingerprint.

Cite this