Author: Stenglein, Mark D.; Sanders, Chris; Kistler, Amy L.; Ruby, J. Graham; Franco, Jessica Y.; Reavill, Drury R.; Dunker, Freeland; DeRisi, Joseph L.
Title: Identification, Characterization, and In Vitro Culture of Highly Divergent Arenaviruses from Boa Constrictors and Annulated Tree Boas: Candidate Etiological Agents for Snake Inclusion Body Disease Document date: 2012_8_14
ID: vkhg20he_36
Snippet: Quantitative PCR. Quantitative PCRs to monitor viral RNA levels in tissues and in culture replication experiments were performed on an LC480 instrument (Roche). RNA was extracted from tissue as described above or from tissue culture supernatant using the ZR viral RNA kit (Zymo Research). Reverse transcription reactions were performed as described for library preparation but used random hexamer priming. qPCR mixtures contained 1Ï« LC480 Sybr Green.....
Document: Quantitative PCR. Quantitative PCRs to monitor viral RNA levels in tissues and in culture replication experiments were performed on an LC480 instrument (Roche). RNA was extracted from tissue as described above or from tissue culture supernatant using the ZR viral RNA kit (Zymo Research). Reverse transcription reactions were performed as described for library preparation but used random hexamer priming. qPCR mixtures contained 1Ï« LC480 Sybr Green master mix (Roche), 0.1 M (each) primer (listed in Table S1 in the supplemental material), and 5 l of 1:20-diluted cDNA.
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