Author: Girardi, Erika; Chane-Woon-Ming, Béatrice; Messmer, Mélanie; Kaukinen, Pasi; Pfeffer, Sébastien
Title: Identification of RNase L-Dependent, 3'-End-Modified, Viral Small RNAs in Sindbis Virus-Infected Mammalian Cells Document date: 2013_11_19
ID: v6uc0ijw_42
Snippet: Plasmid carrying wild-type (WT) and green fluorescent protein (GFP)-SINV genomic sequence (kindly provided by M. C. Saleh) was linearized with XhoI and used as a substrate for in vitro transcription using mMESSAGE mMACHINE capped RNA transcription kit (Ambion) following the manufacturer's instructions. Sindbis viral stock was prepared in BHK21 hamster kidney cells, and titers were measured by plaque assay. HEK293 and Vero cells were maintained in.....
Document: Plasmid carrying wild-type (WT) and green fluorescent protein (GFP)-SINV genomic sequence (kindly provided by M. C. Saleh) was linearized with XhoI and used as a substrate for in vitro transcription using mMESSAGE mMACHINE capped RNA transcription kit (Ambion) following the manufacturer's instructions. Sindbis viral stock was prepared in BHK21 hamster kidney cells, and titers were measured by plaque assay. HEK293 and Vero cells were maintained in Dulbecco's modified Eagle medium (DMEM) (Gibco) supplemented with 10% fetal bovine serum (FBS) (Clontech) in a humidified atmosphere of 5% CO 2 at 37°C. Cells were infected with SINV at an MOI of 0.01. Samples were harvested at 0, 4, 8, 12, 16, 24, and 48 h postinfection (hpi).
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