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Genetics

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JoVE Journal Genetics
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

Article DOI: 10.3791/59414-v • 08:18 min • April 24th, 2019
April 24th, 2019 •
1, 1
1Department of Animal Science, University of Nebraska-Lincoln

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Summary
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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.

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Single Molecule Fluorescent In Situ Hybridization SM-FISH Quantification Localization MRNAs Murine Oocytes PCR Variation Kelsey Timme Graduate Student Material Preparation Female Mice Abdominal Cavity Female Reproductive Tract Ovary Removal Oviduct Cut Holding Medium Ampullae Tear Cumulus Cell Oocyte Complexes Pipette Transfer Hyaluronidase Treatment
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