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Combined DNA-RNA Fluorescent In situ Hybridization FISH to Study X Chromosome Inactivation in Differentiated Female Mouse Embryonic Stem Cells
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Simultaneous RNA-DNA FISH.

Lan-Tian Lai1, Zhenyu Meng2, Fangwei Shao3

  • 1School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore, 637551, Singapore.

Methods in Molecular Biology (Clifton, N.J.)
|January 2, 2016
PubMed
Summary
This summary is machine-generated.

This study introduces a new RNA-DNA FISH method for studying long non-coding RNAs (lncRNAs). The improved protocol preserves fragile RNA signals during DNA denaturation, enabling simultaneous detection of RNA and DNA.

Keywords:
Amino-labeled probesFluorescence in situ hybridization (FISH)Simultaneous RNA–DNA FISHSingle-molecule RNA FISHlncRNA

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Simultaneous RNA-DNA FISH is valuable for analyzing RNA and DNA localization and quantity within cells.
  • Standard FISH protocols often fail due to RNA signal degradation during DNA denaturation.

Purpose of the Study:

  • To develop a robust and simple protocol for simultaneous RNA-DNA FISH.
  • To overcome the limitations of existing methods in preserving RNA integrity.

Main Methods:

  • Utilized amino-labeled nucleic acid probes for RNA FISH.
  • Integrated RNA FISH with DNA FISH under optimized conditions.

Main Results:

  • Developed a protocol that successfully protects RNA signals during DNA denaturation.
  • Achieved simultaneous detection of single RNA molecules and DNA.

Conclusions:

  • The new RNA-DNA FISH protocol is effective for studying lncRNAs.
  • This method offers a reliable approach for simultaneous RNA and DNA detection in cellular studies.