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En alternativ kultur metode til at opretholde genomisk Hypomethylation af mus embryonale stamceller ved hjælp af MEK hæmmer PD0325901 og C-Vitamin
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
An Alternative Culture Method to Maintain Genomic Hypomethylation of Mouse Embryonic Stem Cells Using MEK Inhibitor PD0325901 and Vitamin C
DOI:

11:53 min

June 01, 2018

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Chapters

  • 00:04Title
  • 00:57Grow Mouse ES Cells on Gelatin-coated Dishes
  • 02:48Extract DNA from Cells and Enzymatic Digestion of DNA into Single Nucleosides
  • 05:29Analyze 5mC and 5hmC Using UHPLC-MS/MS
  • 08:34Results: Methylation Status of Mouse Embryonic Stem Cells
  • 10:17Conclusion

Summary

Automatic Translation

Vi beskrev i detaljer to kemiske-baserede protokoller for dyrkning mus embryonale stamceller. Denne nye metode udnytter synergistisk mekanismer til at fremme Tet-medierede oxidation (af C-vitamin) og undertrykke de novo syntesen af 5-methylcytosine (af PD0325901) for at opretholde DNA hypomethylation og pluripotency mus embryonale stamceller.

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