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Isolation of Giant Lampbrush Chromosomes from Living Oocytes of Frogs and Salamanders
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Genetics
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JoVE Journal Genetics
Isolation of Giant Lampbrush Chromosomes from Living Oocytes of Frogs and Salamanders

Isolation of Giant Lampbrush Chromosomes from Living Oocytes of Frogs and Salamanders

DOI:
10.3791/54103-v

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10:07 min

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December 05, 2016

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,
1Department of Embryology,Carnegie Institution for Science

Chapters

  • 00:05Title
  • 01:11Isolation of Germinal Vesicles
  • 02:53Removal of Frog Nuclear Envelopes
  • 04:25Removal of Salamander Nuclear Envelopes
  • 05:17Observation of Lampbrush Chromosomes and Centrifugation of Nuclear Contents
  • 06:33Fixing the Nuclear Contents
  • 07:30Results: Visualization of Axolotl and Frog LBCs
  • 09:14Conclusion

Summary

Automatic Translation

Automatic Translation

We present simple techniques for isolating giant transcriptionally active lampbrush chromosomes from living oocytes of frogs and salamanders. We describe how to observe these chromosomes "alive" by phase contrast or differential interference contrast, and how to fix them for fluorescent in situ hybridization or immunofluorescent staining.

Tags

Keywords: Giant Lampbrush ChromosomesAmphibian OocyteGerminal VesicleNuclear OrganellesTranscriptionSplicingRNA ExportImmunofluorescenceIn Situ HybridizationFrogSalamanderOocyte Culture MediumGV Isolation MediumDispersal MediumNuclear Envelope

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