Author: Mateo, Roberto; Nagamine, Claude M.; Kirkegaard, Karla
Title: Suppression of Drug Resistance in Dengue Virus Document date: 2015_12_15
ID: 6bx2nrui_36
Snippet: Determination of proportion of AflII-marked and unmarked genomes. Supernatants from infected cells were used to extract viral RNA as described by Mateo et al. (30) . Total RNA was used as a template for bulk RT-PCR with a SuperScript III one-step RT-PCR system with platinum Taq (Invitrogen, Grand Island, NY) and the following primers: forward, 5= CGTGGACCGACAAAGACAGATTCTT 3=; reverse, 5= CTTGTCTGC TGACGATCATGTGTG 3=. The resulting amplicon was pu.....
Document: Determination of proportion of AflII-marked and unmarked genomes. Supernatants from infected cells were used to extract viral RNA as described by Mateo et al. (30) . Total RNA was used as a template for bulk RT-PCR with a SuperScript III one-step RT-PCR system with platinum Taq (Invitrogen, Grand Island, NY) and the following primers: forward, 5= CGTGGACCGACAAAGACAGATTCTT 3=; reverse, 5= CTTGTCTGC TGACGATCATGTGTG 3=. The resulting amplicon was purified using the NucleoSpin extract II and PCR cleanup kit (Macherey-Nagel, Deer Park, NY) and digested with AflII. The resulting products were run in 1% agarose-Tris-borate-EDTA (TBE) gels and quantified using AlphaImager EP software (AlphaInnotech, San Jose, CA).
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