Selected article for: "RNA polymerase and viral polymerase"
Author: Van Nguyen, Dung; Van Nguyen, Cuong; Bonsall, David; Ngo, Tue Tri; Carrique-Mas, Juan; Pham, Anh Hong; Bryant, Juliet E.; Thwaites, Guy; Baker, Stephen; Woolhouse, Mark; Simmonds, Peter
Title: Detection and Characterization of Homologues of Human Hepatitis Viruses and Pegiviruses in Rodents and Bats in Vietnam
  • Document date: 2018_2_28
    • ID: 0jtfc271_13
    Snippet: In order to minimize contamination, PCR mastermix preparation, and the addition of templates were carried under separated laminar flow hoods and lab spaces. All of the surfaces, tubes, and equipment were UV irradiated between each PCR. Reverse transcription using SuperScript III reverse transcriptase (Invitrogen, Paisley, UK) was performed according to the manufacturer's instruction. Synthesized cDNA was screened for hepaciviruses and pegiviruses.....
    Document: In order to minimize contamination, PCR mastermix preparation, and the addition of templates were carried under separated laminar flow hoods and lab spaces. All of the surfaces, tubes, and equipment were UV irradiated between each PCR. Reverse transcription using SuperScript III reverse transcriptase (Invitrogen, Paisley, UK) was performed according to the manufacturer's instruction. Synthesized cDNA was screened for hepaciviruses and pegiviruses using a semi-nested PCR protocol with broad spectrum degenerate primers, which can detect all known hepaciviruses and pegiviruses. Amplification conditions (using GoTaq from Promega, Southampton, UK) included 95 • C for 3 min, and 30 cycles of denaturation (94 • C, 30 s), annealing (55 • C, 30 s) and elongation (72 • C, 30 s). Similarly, HEV was screened using broadly reactive primers targeting viral RNA-dependent RNA polymerase as described in Drexler et al., 2012 [11] .

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