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Ovation® Ultralow Library Systems
The Ovation Ultralow Library Systems provide a simple, fast and scalable solution for producing libraries used in next-generation sequencing starting with 1 to 100 ng of DNA. The library construction workflow is suitable for a wide range of sequencing applications, including RNA-Seq, Digital Gene Expression (DGE), genomic DNA sequencing, target capture, amplicon sequencing, ChIP-Seq and more.
As shown in Figure 1, the streamlined workflow consists of four main steps: fragmentation of either genomic DNA or double-stranded cDNA, end repair to generate blunt ends, adaptor ligation for multiplexing or no multiplexing and PCR amplification to produce the final library. The entire workflow, including fragmentation, can be completed in as little as four hours and yields DNA libraries ready for cluster formation and either single read or paired-end sequencing.
The Ovation Ultralow Library Systems have been designed for seamless integration with NuGEN's Ovation Prokaryotic RNA-Seq System (Part No. 9030), Ovation RNA-Seq System V2 (Part No. 7102), Ovation RNA-Seq FFPE System (Part No. 7150) and for DNA sequencing applications with low abundance samples that can input directly into library construction without the need for pre-amplification. The Ovation Ultralow Library System (Part No. 0303) contains reagents for production of non-barcoded libraries, while the Ovation Ultralow IL Multiplex System 1-8 (Part No. 0304) and Ovation Ultralow IL Multiplex System 9-16 (Part No. 0305) each provide eight unique barcoded adaptors for multiplex sequencing. In combination these latter two kits enable up to 16-plex sequencing. The Ovation Ultralow DR Multiplex System 1-8 (Part No. 0330) and Ovation Ultralow DR Multiplex System 9-16 (Part No. 0331) offer the same level of multiplexing but use a dedicated read (DR) barcode design. Lastly, for higher levels of multiplexing the Ovation Ultralow DR Multiplex System 1-96 (Part No. 0329) provides 96 independent DR barcoded adaptors.
Figure 1. The Ovation Ultralow Library System Workflow
Figure 2. Sequencing coverage with E. coli genomic DNA
Sixteen independent sequencing libraries were constructed using 1.0 ng of E. coli genomic DNA with the Ovation Ultralow IL Multiplex System 1-8 (Part No. 0304) and sequenced on the Illumina Genome Analyzer IIx using 40 base-pair single reads. The distribution of reads from each sample is plotted to determine the depth of coverage across the E. coli genome. One million four hundred thousand reads were randomly sub-sampled from each data set to generate the above plots. The grey track represents the theoretical read distribution with randomly chosen 40 base-pair sequences mapped to the E. coli genome. The other overlapping tracks are from the experimental libraries.