Journal
/
Genética
/
Use of Single Molecule Fluorescent In Situ Hybridization (SM-FISH) to Quantify and Localize mRNAs in Murine Oocytes
JoVE Journal
Genética
É necessária uma assinatura da JoVE para visualizar este conteúdo.  Faça login ou comece sua avaliação gratuita.
JoVE Journal Genética
Use of Single Molecule Fluorescent In Situ Hybridization (SM-FISH) to Quantify and Localize mRNAs in Murine Oocytes

Use of Single Molecule Fluorescent In Situ Hybridization (SM-FISH) to Quantify and Localize mRNAs in Murine Oocytes

DOI:
10.3791/59414-v

•

08:18 min

•

April 24, 2019

•

,
1Department of Animal Science,University of Nebraska-Lincoln

Capítulos

  • 00:04Título
  • 00:38Collection of Ovulated Oocytes from Female Mice
  • 02:06SM-FISH Staining of Oocytes
  • 04:21Image Processing
  • 06:21Results: mRNA Quantification Results using the Spot Finding and Tracking Program
  • 07:45Conclusion

Summary

Tadução automática

Tadução automática

To reproducibly count the numbers of mRNAs in individual oocytes, single molecule RNA fluorescence in situ hybridization (RNA-FISH) was optimized for non-adherent cells. Oocytes were collected, hybridized with the transcript specific probes, and quantified using an image quantification software.

Tags

Single Molecule Fluorescent In Situ Hybridization (SM-FISH)MRNA QuantificationMRNA LocalizationMurine OocytesNanogPou5f1DapBOocyte PreparationFixationPermeabilizationProtease TreatmentProbe Hybridization

Article

Embed

ADICIONAR À PLAYLIST

Usage Statistics

Vídeos Relacionados

Optogenetic Random Mutagenesis Using Histone-miniSOG in C. elegans

Detection of Copy Number Alterations Using Single Cell Sequencing

Detection of Inter-chromosomal Stable Aberrations by Multiple Fluorescence In Situ Hybridization (mFISH) and Spectral Karyotyping (SKY) in Irradiated Mice

Generation of Fluorescent Protein Fusions in Candida Species

Single Molecule Analysis of Laser Localized Psoralen Adducts

High Resolution Fluorescent In Situ Hybridization in Drosophila Embryos and Tissues Using Tyramide Signal Amplification

Visualization of Microbiota in Tick Guts by Whole-mount In Situ Hybridization

Single Molecule Fluorescence In Situ Hybridization (smFISH) Analysis in Budding Yeast Vegetative Growth and Meiosis

Quantitative Analysis of Alternative Pre-mRNA Splicing in Mouse Brain Sections Using RNA In Situ Hybridization Assay

Tissue-specific miRNA Expression Profiling in Mouse Heart Sections Using In Situ Hybridization

Read Article