Author: Gebhardt, Jordan T; Woodworth, Jason C; Jones, Cassandra K; Tokach, Mike D; Gauger, Philip C; Main, Rodger G; Zhang, Jianqiang; Chen, Qi; DeRouchey, Joel M; Goodband, Robert D; Stark, Charles R; Bergstrom, Jon R; Bai, Jianfa; Dritz, Steve S
Title: Determining the impact of commercial feed additives as potential porcine epidemic diarrhea virus mitigation strategies as determined by polymerase chain reaction analysis and bioassay() Document date: 2018_8_20
ID: 6rlbiukh_14
Snippet: After collection of d 42 post-laboratory inoculation aliquots, qRT-PCR was conducted on designated preserved aliquots at Kansas State University Veterinary Diagnostic Laboratory Molecular Diagnostics Laboratory. Fifty microliters of supernatant from each sample was loaded into a deep-well plate and extracted using a Kingfisher 96 magnetic particle processor (Fisher Scientific, Pittsburg, PA) and the MagMAX-96 Viral RNA Isolation kit (Life Technol.....
Document: After collection of d 42 post-laboratory inoculation aliquots, qRT-PCR was conducted on designated preserved aliquots at Kansas State University Veterinary Diagnostic Laboratory Molecular Diagnostics Laboratory. Fifty microliters of supernatant from each sample was loaded into a deep-well plate and extracted using a Kingfisher 96 magnetic particle processor (Fisher Scientific, Pittsburg, PA) and the MagMAX-96 Viral RNA Isolation kit (Life Technologies, Grand Island, NY) according to the manufacturer's instructions with one modification, reducing the final elution volume to 60 µL. One negative extraction control consisting of all reagents except the sample was included in each extraction. The extracted RNA was frozen at −20 °C until assayed by qRT-PCR. Analyzed values represent cycle threshold (Ct) at which virus was detected. A greater Ct value indicates more cycles must proceed until viral genetic material is detected, thus lower quantities of genetic material are present in the original sample.
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