Encore™ NGS Multiplex System I

The Encore NGS Library Systems provides a simple, fast, and scalable solution for producing DNA libraries used in a wide range of sequencing applications including RNA-Seq, Digital Gene Expression (DGE), genomic DNA/exome sequencing, amplicon sequencing, ChIP-Seq and more.

As shown in Figure 1, the streamlined workflow consists of four main steps: (1) Fragmentation of either genomic DNA or double-stranded cDNA to produce fragments with a tight size distribution; (2) End repair to generate blunt ends; (3) Adaptor ligation for multiplexing or no multiplexing; and (4) PCR amplification to produce the final library. The entire workflow including fragmentation can be completed in as little as three hours, and yields DNA libraries ready for cluster formation and either single read or paired-end sequencing.

The Encore NGS Library Systems have been designed for seamless integration with both the Ovation RNA-Seq System and Ovation 3'-DGE System to enable a complete end-to-end solution for transcriptome library construction using unfractionated total RNA samples in 9-10 hours.

Table 1 shows a clear segregation of libraries using the index tags, and even distribution of reads derived from libraries containing each index. There is no preferential cluster formation or sequencing from any of the indexed libraries. The theoretical distribution of indexed Reads for an 8-plex run is 12.5%.

Together, the portfolio of integrated NGS products from NuGEN provides a complete, flexible and robust solution to all your NGS sample preparation needs.

Table 1: Independent DNA libraries were constructed from each of the samples listed below using the Encore NGS Multiplex System I, and equimolar amounts pooled for addition to the Illumina cBot

10 ng of Human MAQC A and B samples were amplified using the Ovation 3'-DGE System and 100 ng Genomic DNA was isolated from E. coli. Independent DNA libraries were constructed from each of these samples using the Encore Multiplex System I, and equimolar amounts pooled for addition to the Illumina cBot. 8-Plex single-read sequencing was done in a single lane with 40 base-pair reads on the Illumina Genome Analyzer IIx platform, and resulting reads aligned to either the E. coli reference genome or the human hg18 reference.

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