Abstract
Long-read RNA sequencing allows for the precise characterization of full-length transcripts, which makes it an indispensable tool in transcriptomics. The human cytomegalovirus (HCMV) genome has been first sequenced in 1989 and although short-read sequencing studies have uncovered much of the complexity of its transcriptome, only few of its transcripts have been fully annotated. We hereby present a long-read RNA sequencing dataset of HCMV infected human lung fibroblast cells sequenced by the Pacific Biosciences RSII platform. Seven SMRT cells were sequenced using oligo(dT) primers to reverse transcribe poly(A)-selected RNA molecules and one library was prepared using random primers for the reverse transcription of the rRNA-depleted sample. Our dataset contains 122,636 human and 33,086 viral (HMCV strain Towne) reads. The described data include raw and processed sequencing files, and combined with other datasets, they can be used to validate transcriptome analysis tools, to compare library preparation methods, to test base calling algorithms or to identify genetic variants.
Highlights
Background & SummaryLong-read sequencing surveys of eukaryotic transcriptomes have demonstrated the potential of this new technology in identifying novel transcripts and characterizing transcript isoforms[1,2,3]
More transcriptomic data generated by long-read sequencing would facilitate the development of analysis tools needed to evaluate such data
A recent Illumina-based short-read sequencing study has shown that the Human cytomegalovirus (HCMV) transcriptome is more complex than it had been recognized previously[8]
Summary
Long-read sequencing surveys of eukaryotic transcriptomes have demonstrated the potential of this new technology in identifying novel transcripts and characterizing transcript isoforms[1,2,3]. Seven sequencing runs were carried out using oligo(dT) selection methods, to analyse the polyadenylated fraction of transcripts and one library was prepared by random primer amplification to capture non-polyadenylated transcripts as well. Our aim with these experiments was to assess the utility of Pacific Biosciences isoform sequencing (Iso-Seq) sequencing in the transcriptome profiling of HCMV, to identify novel viral transcripts and to complement the already existing viral transcriptome[9]. As the pooled samples contained RNA from early post infection time points, when host transcription has not yet been disrupted by the virus, most of the reads (122,636 reads) aligned to the human genome.
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