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Use of Bisection to Reduce Mitochondrial DNA in the Bovine Oocyte
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发育生物学
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JoVE 杂志 发育生物学
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

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06:15 min

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July 06, 2022

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, , , , , ,
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

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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

BisectionMitochondrial DNABovine OocyteCloningHeteroplasmyCentrifugationZonae PellucidaePronaseCBT20

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