METTL3 and DNA staining of SN&NSN GV oocytes

Published: 28 December 2022| Version 1 | DOI: 10.17632/dhskwr3r5f.1
Contributor:
Yao Huan

Description

There are both SN (Surrounded nucleus) and NSN (Non surrounded nucleus) GV oocytes in the cumulus cell-oocyte complex of both control and Mettl3 cKO mice after PMSG injection.

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Steps to reproduce

Oocytes were fixed in 4% PFA for 30 min at room temperature (RT) followed by permeabilization in 0.25% Triton X-100 for 15 min before blocking in 3% BSA for 1 h at RT. and incubated with primary antibodies in 3% BSA overnight at 4℃. The embryos were washed 3 times for 5 min each in washing solution (0.1% Tween-20 in PBS) for 1 h at RT and incubated with secondary antibodies (Alexa Fluor conjugated; 1:500 in washing solution) for 1 h at RT. After being washed thoroughly three times in washing solution, embryos were stained with DAPI (1:1000 in washing solution) for 5 min and subjected to take images using a confocal microscope.

Institutions

Beijing Institute of Genomics Chinese Academy of Sciences

Categories

Fluorescence Labeling

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