Benchmarking Sequence Technologies for Virome Sequencing
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ABSTRACT:
PROVIDER: PRJEB56639 | ENA |
REPOSITORIES: ENA
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2012-06-09 | E-GEOD-34478 | biostudies-arrayexpress
Project description:Next-generation sequencing has emerged as a promising platform for whole genome transcriptome profiling. The microarray technology has in contrast a long track record, but may suffer from issues related to background fluorescence and non-specific hybridization. The two technologies have been compared for eukaryotic transcriptomes in various studies. However, for prokaryotes information on comparative performance of the two technologies is sparse. We thus compared the two technologies for Streptococcus thermophilus. With just single experiments there was a strong correlation between the technologies. We compared our in house spotted DNA microarray platform with the same RNA samples sequenced externally. The aim was to, without replicate arrays or sequencing, to determine if the two technologies correlate well.
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Project description:Virome sequencing of human stool
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Project description:Next-generation sequencing has emerged as a promising platform for whole genome transcriptome profiling. The microarray technology has in contrast a long track record, but may suffer from issues related to background fluorescence and non-specific hybridization. The two technologies have been compared for eukaryotic transcriptomes in various studies. However, for prokaryotes information on comparative performance of the two technologies is sparse. We thus compared the two technologies for Streptococcus thermophilus. With just single experiments there was a strong correlation between the technologies.
2012-06-10 | GSE34461 | GEO
Project description:Next-generation sequencing has emerged as a promising platform for whole genome transcriptome profiling. The microarray technology has in contrast a long track record, but may suffer from issues related to background fluorescence and non-specific hybridization. The two technologies have been compared for eukaryotic transcriptomes in various studies. However, for prokaryotes information on comparative performance of the two technologies is sparse. We thus compared the two technologies for Streptococcus thermophilus. With just single experiments there was a strong correlation between the technologies.
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Project description:RNA-sequencing has become the gold standard for whole-transcriptome gene expression quantification. Multiple algorithms have been developed to derive gene counts from sequencing reads. While a number of benchmarking studies have been conducted, the question remains how individual methods perform at accurately quantifying gene expression levels from RNA-sequencing reads. We performed an independent benchmarking study using RNA-sequencing data from the well established MAQCA and MAQCB reference samples. RNA-sequencing reads were processed using five popular workflows (Tophat-HTSeq, Tophat-Cufflinks, STAR-HTSeq, Kallisto and Salmon) and resulting gene expression measurements were compared to expression data generated by wet-lab validated qPCR assays for all protein coding genes. All methods showed high gene expression rank correlations with qPCR data. When comparing gene expression fold changes between MAQCA and MAQCB samples, about 85% of the genes showed consistent results between RNA-sequencing and qPCR data. Of note, each method revealed a small but specific set of genes with inconsistent expression measurements. A significant proportion of these method-specific inconsistent genes were reproducibly identified in independent datasets. These genes were typically smaller, had fewer exons and were lower expressed compared to genes with consistent expression measurements. We propose that careful validation is warranted when evaluating RNA-seq based expression profiles for this specific set of genes.
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