Selected article for: "genome assembly and Illumina protocol"

Author: Alexander M. Price; Katharina E. Hayer; Daniel P. Depledge; Angus C. Wilson; Matthew D. Weitzman
Title: Novel splicing and open reading frames revealed by long-read direct RNA sequencing of adenovirus transcripts
  • Document date: 2019_12_13
  • ID: 2fcl6f4i_15
    Snippet: Adenovirus serotype 5 (Ad5) was originally purchased from ATCC. All viruses were 307 expanded on HEK293 cells, purified using two sequential rounds of ultracentrifugation in CsCl 308 gradients, and stored in 40% v/v glycerol at -20 °C (short term) or -80 °C (long term). Viral stock 309 titer was determined on HEK293 cells by plaque assay, and all subsequent infections were 310 The copyright holder for this preprint (which was not peer-reviewed).....
    Document: Adenovirus serotype 5 (Ad5) was originally purchased from ATCC. All viruses were 307 expanded on HEK293 cells, purified using two sequential rounds of ultracentrifugation in CsCl 308 gradients, and stored in 40% v/v glycerol at -20 °C (short term) or -80 °C (long term). Viral stock 309 titer was determined on HEK293 cells by plaque assay, and all subsequent infections were 310 The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2019.12.13.876037 doi: bioRxiv preprint Total RNA from three biological replicates of Control knockdown or three biological 325 replicates of METTL3-knockdown A549 cells infected with Ad5 for 24 hours were sent to Genewiz 326 for preparation into strand-specific RNA-Seq libraries. Libraries were then run spread over three 327 lanes of an Illumina HiSeq 2500 using a 150bp paired-end protocol. Raw reads were mapped to 328 the GRCh37/hg19 genome assembly and the Ad5 genome using the RNA-seq aligner GSNAP 329

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