A Video Protocol of Retroviral Infection in Primary Intestinal Organoid Culture (original) (raw)
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09:18 min
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August 11th, 2014
DOI :
August 11th, 2014
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Amanda Andersson-Rolf1,2, Juergen Fink1,2, Roxana C. Mustata2, Bon-Kyoung Koo1,2
1Department of Genetics, University of Cambridge, 2Wellcome Trust - Medical Research Council Stem Cell Institute, University of Cambridge
0:05
Title
1:17
Crypt Isolation
3:39
Virus Production
5:15
Retroviral Transduction
6:34
Spinoculation and Plating
8:01
Results: Representative Organoid Images
8:49
Conclusion
This protocol explains primary Lgr5-positve organoid culture and the subsequent performance of retroviral transduction. This enables Cre-inducible overexpression or knockdown of the delivered transgene and allows functional studies to be carried out in the novel in vitro organotypicmodel system_._
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