Author: Malboeuf, Christine M.; Yang, Xiao; Charlebois, Patrick; Qu, James; Berlin, Aaron M.; Casali, Monica; Pesko, Kendra N.; Boutwell, Christian L.; DeVincenzo, John P.; Ebel, Gregory D.; Allen, Todd M.; Zody, Michael C.; Henn, Matthew R.; Levin, Joshua Z.
Title: Complete viral RNA genome sequencing of ultra-low copy samples by sequence-independent amplification Document date: 2012_9_8
ID: s76c5ebd_8
Snippet: WNV was produced from an infectious cDNA clone according to methods described elsewhere (29) . Briefly, hamster BHK-21 cells (ATCC) were transfected (transMessenger, Qiagen) with RNA transcribed in vitro from pFLWNV (29) , using the mMessage mMachine T7 kit (Life Technologies, Carlsbad, CA). Cells were inspected daily and virus harvested after approximately 3 days of replication. Virus-containing tissue culture supernatant was supplemented with 2.....
Document: WNV was produced from an infectious cDNA clone according to methods described elsewhere (29) . Briefly, hamster BHK-21 cells (ATCC) were transfected (transMessenger, Qiagen) with RNA transcribed in vitro from pFLWNV (29) , using the mMessage mMachine T7 kit (Life Technologies, Carlsbad, CA). Cells were inspected daily and virus harvested after approximately 3 days of replication. Virus-containing tissue culture supernatant was supplemented with 20% FBS, stored at À80 C in 1 ml aliquots and used without subsequent passage.
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