Author: Peña, José; Chen-Harris, Haiyin; Allen, Jonathan E.; Hwang, Mona; Elsheikh, Maher; Mabery, Shalini; Bielefeldt-Ohmann, Helle; Zemla, Adam T.; Bowen, Richard A.; Borucki, Monica K.
Title: Sendai virus intra-host population dynamics and host immunocompetence influence viral virulence during in vivo passage Document date: 2016_4_9
ID: z7f720dj_23
Snippet: Sequencing control plasmids were generated to determine the error rate of the PCR and Illumina sequencing as described previously in Chen-Harris et al. (2013) . To gauge sensitivity of the protocol, dilutions of control plasmids containing inserts from SeV, rabies virus (RABV) and bovine coronavirus (BCoV) were combined at following ratios 100,000 SeV: 1,000 RABV: 10 BCoV. Plasmid copy number for each plasmid control was estimated via spectrophot.....
Document: Sequencing control plasmids were generated to determine the error rate of the PCR and Illumina sequencing as described previously in Chen-Harris et al. (2013) . To gauge sensitivity of the protocol, dilutions of control plasmids containing inserts from SeV, rabies virus (RABV) and bovine coronavirus (BCoV) were combined at following ratios 100,000 SeV: 1,000 RABV: 10 BCoV. Plasmid copy number for each plasmid control was estimated via spectrophotometer and Taqman assay (Borucki et al. 2013a,b) Viral inserts present in the plasmid mixtures were amplified using M13 primers and Phusion PCR as described previously. This process was done in replicate to generate two independent samples for use as Illumina sequencing controls.
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