Abstract
The first complete coding sequence of the Venezuelan equine encephalitis virus IE, isolated from a Costa Rican mare with severe encephalitis, was confirmed by histological and viral whole-genome analyses. The isolated virus grouped in the Pacific cluster.
Highlights
The supernatant of a single passage on Vero E6 cells infected with Venezuelan equine encephalitis virus (VEEV) was clarified by centrifugation and extracted with TRIzol LS (Invitrogen, USA) [10]
Quantity were determined using a NanoDrop spectrophotometer (Thermo Fisher, USA) and a Quantus fluorometer (Promega, USA). cDNA synthesis was conducted using random hexamers and the SuperScript III kit per the manufacturer’s instructions (Invitrogen). cDNA was treated with Klenow DNA polymerase (Applied Biosystems, USA) and RNase H (Invitrogen) at 37°C for 1 h 10 min at 75°C
Quantity, and fragment size distribution of nucleic acids were evaluated with a NanoDrop spectrophotometer, a Quantus fluorometer (Promega), and a QIAxcel system (Qiagen, USA), respectively
Summary
The supernatant of a single passage on Vero E6 cells infected with VEEV was clarified by centrifugation and extracted with TRIzol LS (Invitrogen, USA) [10]. Quantity were determined using a NanoDrop spectrophotometer (Thermo Fisher, USA) and a Quantus fluorometer (Promega, USA). CDNA synthesis was conducted using random hexamers and the SuperScript III kit per the manufacturer’s instructions (Invitrogen). CDNA was treated with Klenow DNA polymerase (Applied Biosystems, USA) and RNase H (Invitrogen) at 37°C for 1 h 10 min at 75°C.
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