Yellow fever virus
Protocols: Yellow Fever infectious clones (IC) were generated by site-directed mutagenesis (SDM) of either pANCR 17D (IC) or pANCR Asibi IC as a template. QuickChange XLII SDM kit (Strategene) along with mutagenic primer sets were used to generate infectious clones as per the manufacturer's protocol. Stock cDNA YF IC were amplified in E. coli (MC1061) and plasmid harvested by large-scale extraction. Full-length YF RNA was generated by in vitro transcription of purified cDNA using SP6-Scribe standard RNA in vitro transctiption kit (Cellscript) according to the manufacturer's protocol; the reaction was incubated at 37oC for 2.5 hours. The entire reaction was electroporated into 6.7x106 monkey kidney Vero cells. Cell supernatant was collected when infected cells showed 80% cytopathic effect (CPE). RNA from the YF ICs were extracted using QIAamp Viral RNA mini kit (Qiagen), following the manufacturer's guidelines. NGS libraries were constructed using TruSeq RNA v2 kit as recommended by the manufacturer.
ENA-FIRST-PUBLIC
2018-10-16T17:02:47Z
ENA-LAST-UPDATE
2018-03-20T16:28:12Z
External Id
SAMEA104708359
INSDC center name
Department of Microbiology & Immunology, University of Texas Medical Branch
INSDC first public
2018-10-16T17:02:47Z
INSDC last update
2018-03-20T16:28:12Z
INSDC status
public
Submitter Id
E-MTAB-6614:Sample 12
broker name
ArrayExpress
genotype
wild type genotype
growth condition
grown in monkey kidney Vero cells
sample name
E-MTAB-6614:Sample 12
scientific_name
Yellow fever virus
strain
Asibi