Ovation® SP Ultralow Library Systems
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The Ovation SP Ultralow Library and DR Multiplex Systems provide automated, fast and scalable solutions for producing both non-multiplexed and multiplexed libraries starting with as little as 1.0 ng of DNA. The resulting libraries are suitable for a wide range of sequencing applications on Illumina sequencing-by-synthesis platforms, including RNA-Seq, Digital Gene Expression (DGE), genomic DNA/exome sequencing, amplicon sequencing, ChIP-Seq and more.
As shown in Figure 1 (below), the streamlined workflow consists of six steps:
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Fragmentation of either genomic DNA or double-stranded cDNA to produce assay template
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Addition of template and reagents to the Mondrian SP Universal Cartridge
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Hands-free automation of the following assay steps on the Mondrian SP Workstation:
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Sample concentration
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End repair
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Sample purification
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Adaptor ligation (with optional sample multiplexing)
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Sample purification
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PCR enrichment of the purified library
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Enriched library purification and quantification
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Cluster formation and sequencing
The figure below displays the distribution of average read depth vs number of bases for 4 different E. coli gDNA libraries (ranging from 5 ng to 0.1 ng starting material) and a control of 2.4 million random E. coli fragments. The average read depth of all the libraries is between 16 and 18x (regardless of the amount of starting material in the assay), illustrating that Ovation SP Ultralow Library Systems generate high quality libraries from as little as 1.0 ng of starting material.
The Ovation SP 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 to the library construction without the need for pre-amplification.
The Ovation SP Ultralow Library System (Part No. 8030) contains reagents for production of non-barcoded libraries, while the Ovation SP Ultralow DR Multiplex System 1-8 (Part No. 8033) and Ovation SP Ultralow DR Multiplex System 9-16 (Part No. 8034) provide eight unique barcoded adaptors for multiplex sequencing. In combination these latter two kits enable up to 16-plex sequencing using a dedicated read (DR) barcode design.
October 26, 2012 presentation by Dr. Tomas Babak describing the analysis of human whole genome sequence data generated from a small DNA sample (50 nanograms) using the Ovation Ultralow SP DNA Library System. The library was prepared on the Mondrian SP digital microfluidics system.