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SRX5774957: Cell fusing agent virus isolate KPP, RNA produced
1 ILLUMINA (NextSeq 500) run: 2.9M spots, 423M bases, 188.1Mb downloads

Design: For preparation of cDNA libraries RNA was extracted using QIAamp Viral RNA Mini Kit (Qiagen) according to manufacturers instructions and eluted in 60 L of AE buffer. RNA was subsequently treated with TURBO DNase (Ambion), and cDNA was produced using M-MLV reverse transcriptase (Invitrogen) following manufacturers instructions. cDNA was incubated for 2h at 16C with E. coli DNA ligase (New England BioLabs), E. coli DNA polymerase I (New England BioLabs), and E. coli RNase H (New England BioLabs) for second strand synthesis with Second Strand Synthesis Buffer (New England BioLabs). Purified dsDNA was used for library preparation using Nextera XT DNA Kit (Illumina) according to manufacturers instructions followed by cDNA quality check by Bioanalyzer DNA 1000 kit (Agilent). The libraries were combined with other libraries from unrelated projects and loaded onto a NextSeq 500/550 Mid-Output Kit v2 150 cycles flow cell (Illumina)
Submitted by: Institut Pasteur
Study: Cell fusing agent virus sequencing
show Abstracthide Abstract
Aedes aegypti mosquitoes are vectors for such medically important arboviruses as dengue or Zika viruses. However mosquitoes are also naturally infected with other viruses that belong to genus Flavivirus, that can only replicate in insect cells - insect specific flaviviruses. One of the insect specific flaviviruses, cell fusing agent virus (CFAV) had been isolated from the 4th generation of Thai Ae. aegypti colony. The goal of this project was to sequence full genome of CFAV isolated on C6/36 cell line from mosquito homogenates. To omit potential coisolation of other viruses from mosquito homogenates, we prepared an additional CFAV stock directly from CFAV genomic RNA. In addition, we sequenced cDNA from naive C6/36 supernatant to verify its virus content.
Sample:
SAMN11520269 • SRS4708445 • All experiments • All runs
Library:
Name: CFAV-KPP.OK3
Instrument: NextSeq 500
Strategy: RNA-Seq
Source: VIRAL RNA
Selection: RANDOM
Layout: PAIRED
Runs: 1 run, 2.9M spots, 423M bases, 188.1Mb
Run# of Spots# of BasesSizePublished
SRR89960762,930,958423M188.1Mb2019-05-01

ID:
7765460

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