Journal
/
Developmental Biology
/
Use of Bisection to Reduce Mitochondrial DNA in the Bovine Oocyte
JoVE Journal
Developmental Biology
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Developmental Biology
Use of Bisection to Reduce Mitochondrial DNA in the Bovine Oocyte

Use of Bisection to Reduce Mitochondrial DNA in the Bovine Oocyte

DOI:
10.3791/64060-v

•

06:15 min

•

July 06, 2022

•

, , , , , ,
1Department of Animal, Dairy, and Veterinary Sciences,Utah State University, 2Reproductive Sciences, Beckman Center for Conservation Research,San Diego Zoo Wildlife Alliance, 3Genus PLC

Chapters

  • 00:04Introduction
  • 00:47Centrifugation of the Oocytes
  • 01:51Preparation of the Oocyte for Bisection
  • 03:00Bisection of the Oocytes
  • 04:56Results: Successful and Unsuccessful qPCR Results and Boxplot from Mitochondria-Reduced and Mitochondria-Dense Ooplasts
  • 05:39Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we present a protocol to significantly reduce the mitochondrial DNA copy numbers in a bovine oocyte (P < 0.0001). This method utilizes centrifugation and bisection to substantially reduce oocyte mitochondria and may allow for an increased chance of development in the reconstructed interspecies somatic cell nuclear transfer embryos.

Tags

Keywords: BisectionMitochondrial DNABovine OocyteCloningHeteroplasmyCentrifugationZonae PellucidaePronaseCBT20

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/64060?article_type=v

Production and Use of Lentivirus to Selectively Transduce Primary Oligodendrocyte Precursor Cells for In Vitro Myelination Assays
check_url/64060?article_type=v

Use of the TetON System to Study Molecular Mechanisms of Zebrafish Regeneration
check_url/64060?article_type=v

Functional Cloning Using a Xenopus Oocyte Expression System
check_url/64060?article_type=v

Technique to Target Microinjection to the Developing Xenopus Kidney
check_url/64060?article_type=v

Functional Manipulation of Maternal Gene Products Using In Vitro Oocyte Maturation in Zebrafish
check_url/64060?article_type=v

A 5-mC Dot Blot Assay Quantifying the DNA Methylation Level of Chondrocyte Dedifferentiation In Vitro
check_url/64060?article_type=v

Use of Immunolabeling to Analyze Stable, Dynamic, and Nascent Microtubules in the Zebrafish Embryo
check_url/64060?article_type=v

Dissection and Immunofluorescent Staining of Mushroom Body and Photoreceptor Neurons in Adult Drosophila melanogaster Brains
check_url/64060?article_type=v

Application of RNAi and Heat-shock-induced Transcription Factor Expression to Reprogram Germ Cells to Neurons in C. elegans
check_url/64060?article_type=v

Use of Hematopoietic Stem Cell Transplantation to Assess the Origin of Myelodysplastic Syndrome

Read Article