Encore™ NGS Library System I and 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.

As shown in Figure 2, even more convenience and flexibility are possible when using the Prelude Direct Lysis Module. These data illustrate that very similar RNA-Seq results are obtained using direct cell lysates or isolated total RNA. Taken together, the portfolio of integrated NGS products from NuGEN provides a complete, flexible and robust solution to all your NGS sample preparation needs.

Figure 2:

U2000 U937 cells were processed using the Prelude Direct Lysis Module, or total RNA was isolated from U937 cells using the QIAGEN® RNeasy® Mini Kit. Cell lysates or 20 ng of isolated total RNA was amplified using the Ovation RNA-Seq System, and libraries constructed using the Encore NGS Library System I. Single-read sequencing was done with 40 base-pair reads on the Illumina Genome Analyzer IIx platform. Fastq reads were processed with Tophat v1.0.11 and Bowtie 0.12.3 to generate RPKM values (Reads Per Kilobase per Million mapped reads) for 20,420 transcripts. Log, RPKM values are plotted

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