Zum Anzeigen dieser Inhalte ist ein JoVE-Abonnement erforderlich. Melden Sie sich an oder starten Sie Ihre kostenlose Testversion.
Isolation of Giant Lampbrush Chromosomes from Living Oocytes of Frogs and Salamanders
Kapitel
- 00:05Titel
- 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
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.
