Author: Rhein, Bethany A.; Powers, Linda S.; Rogers, Kai; Anantpadma, Manu; Singh, Brajesh K.; Sakurai, Yasuteru; Bair, Thomas; Miller-Hunt, Catherine; Sinn, Patrick; Davey, Robert A.; Monick, Martha M.; Maury, Wendy
Title: Interferon-? Inhibits Ebola Virus Infection Document date: 2015_11_12
ID: 10bu7iwg_69
Snippet: Loading of DsiRNA into exosomes. Dicer-substrate RNAs (DsiRNA) (IDT), 27mer duplex RNAs that have increased potency in RNA interference compared to tradition siRNAs [97] , were loaded into exosomes by electroporation as described [98] . Briefly, exosome pellets were resuspended in electroporation buffer for siRNA loading. The electroporation mixture was prepared by mixing exosomes and DsiRNA in 1:1 (wt/wt) in electroporation buffer. The final con.....
Document: Loading of DsiRNA into exosomes. Dicer-substrate RNAs (DsiRNA) (IDT), 27mer duplex RNAs that have increased potency in RNA interference compared to tradition siRNAs [97] , were loaded into exosomes by electroporation as described [98] . Briefly, exosome pellets were resuspended in electroporation buffer for siRNA loading. The electroporation mixture was prepared by mixing exosomes and DsiRNA in 1:1 (wt/wt) in electroporation buffer. The final concentration of exosomes in the mixture was 250 ng/μL. The mixture was electroporated in a 400 μL volume using 0.4-mm cuvettes at 400 mV and 125 μF capacitance with pulse time of 10-15 ms. Exosomes were aliquoted and frozen at -20°C until use.
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