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. 2015 Apr;12(4):323-5.
doi: 10.1038/nmeth.3313. Epub 2015 Mar 2.

Simultaneous generation of many RNA-seq libraries in a single reaction

Alexander A Shishkin  1 , Georgia Giannoukos  2 , Alper Kucukural  3 , Dawn Ciulla  2 , Michele Busby  2 , Christine Surka  1 , Jenny Chen  4 , Roby P Bhattacharyya  2 , Robert F Rudy  2 , Milesh M Patel  2 , Nathaniel Novod  2 , Deborah T Hung  2 , Andreas Gnirke  2 , Manuel Garber  5 , Mitchell Guttman  1 , Jonathan Livny  2
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Simultaneous generation of many RNA-seq libraries in a single reaction

"V体育平台登录" Alexander A Shishkin et al. Nat Methods. 2015 Apr.

"VSports最新版本" Abstract

Although RNA-seq is a powerful tool, the considerable time and cost associated with library construction has limited its utilization for various applications VSports手机版. RNAtag-Seq, an approach to generate multiple RNA-seq libraries in a single reaction, lowers time and cost per sample, and it produces data on prokaryotic and eukaryotic samples that are comparable to those generated by traditional strand-specific RNA-seq approaches. .

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Figures

Figure 1
Figure 1
Schematic of RNAtag-Seq method. Gray and black lines correspond to RNA and cDNA, respectively. Colored blocks represent unique sequence barcodes. Light green lines and purple bars represent Illumina sequencing adaptors and Illumina index barcodes, respectively.
Figure 2
Figure 2
Differential gene expression analysis using RNAtag-Seq. (a) Heat map of all 3,875 differentially expressed genes (fold change >2, Padj < 0.01 with Padj corresponding to the P value adjusted for multiple testing using the Benjamini-Hochberg procedure) across adult mouse tissues and mouse embryos at developmental stages E11 and E15. (b) Selected Gene Ontology categories and their enrichment for specific tissues: brain and spinal cord samples (top) and eye samples (bottom) relative to all samples. The enrichment is plotted as the −log10 of the enrichment P value. (c) MA plots of 2 ciprofloxacin-susceptible (CipS, left) and 2 ciprofloxacin-resistant (CipR, right) E. coli clinical isolates 30 min after exposure to ciprofloxacin versus untreated. Genes found to be significantly up- and downregulated (greater than threefold, Padj < 0.05) by RNAtag-Seq are colored black. Genes in the SOS regulon are colored red.
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