Flexibility to fit your needs
Scalable batch sizes and continuous loading of multiple
flow cells enable you to adapt and scale the GeneReader NGS System to match your
needs and grow
Images
Better sequence coverage and minimal bias.
A mixture of highly GC-rich Bordetella pertussis gDNA (GC content 67.7%) and highly AT-rich Streptobacillus moniliformis gDNA (GC content 26.3%) was pooled, made into an Illumina-compatible DNA library using the GeneRead Library Prep (I) Kit, and amplified with the GeneRead Library Amp (I) Kit, which contains HiFi Polymerase, or a kit from Supplier K. The amplified libraries were sequenced using the Illumina MiSeq instrument, and fidelity and sequence coverage was analyzed using the Galaxy platform. The GeneRead Library Prep Kit provided greater sequence coverage in GC- and AT-rich areas of DNA, compared to the kit from Supplier K.
Targeted enrichment NGS workflow.
First, extract DNA (the QIAamp DNA Mini Kit or QIAamp DNA FFPE Tissue Kit is recommended), and then use GeneRead DNAseq Gene Panels for targeted exon enrichment. Then construct your NGS library, quantify and quality-control using the GeneRead Library Quantification System, and perform NGS and data analysis using the QIAGEN NGS Data Analysis Web Portal.
High yields of library DNA.
Genomic DNA (1 μg) from E. coli strain DH10B was sheared using a Covaris instrument and made into an Illumina-compatible DNA library using the GeneRead Library Prep (I) Kit or a kit from another supplier. The library was eluted in 18 μl Buffer EB and library concentrations were determined by quantitative PCR.
Low error rates with minimal sequence bias.
A mixture of highly GC-rich Bordetella pertussis gDNA (GC content 67.7%) and highly AT-rich Streptobacillus moniliformis gDNA (GC content 26.3%) was pooled, made into an Illumina-compatible DNA library using the GeneRead Library (I) Core Kit, and amplified with the GeneRead Library (I) Amp Kit, which contains HiFi Polymerase, or a kit from Supplier K. The amplified libraries were sequenced using the Illumina MiSeq instrument, and fidelity and sequence coverage were analyzed using the Galaxy platform. [A] Low error rates and [B] greater cumulated sequence coverage demonstrate that the GeneRead Library (I) Amp Kit provides superior library amplification compared to the kit from Supplier K.
Optimized workflow for Illumina.
The GeneRead Library Prep (I) Kit uses an optimized one-tube protocol, with fewer cleanup steps and optional high-fidelity library amplification. The hands-on time and total time required for preparation of library DNA is significantly reduced compared to the library preparation system from Supplier I.
Optimized workflow for Life Technologies.
The GeneRead Library Prep (L) Kit uses an optimized, one-tube procedure, with fewer cleanup steps and optional high-fidelity library amplification. The hands-on time and total time required for preparation of library DNA is significantly reduced compared to the library preparation system from Supplier L.
PCR-enabled target enrichment of genes of interest.
The principle of the GeneRead DNAseq System is to employ overlapping primer sets across the exonic portions of a gene or genes to maximize target coverage and minimize nonspecific amplification.
Precise size selection.
The GeneRead Size Selection Kit effectively removes adapter dimers and adapter monomers following library preparation. A scaled-up image of the above data showing the correct size distribution of Illumina-compatible library fragments following size selection is shown in inset. FU: Fluorescence units.
Quantification of libraries with concentrations below the detection limit of conventional methods.
The GeneRead Library Quant Array's high sensitivity and broad dynamic range enable the quantification of both NGS-L1 and NGS-L2 libraries. By contrast, the kit from Suppler A (for use with the Agilent 2100 BioAnalyzer) quantified only the NGS-L1 library; with this kit, the NGS-L2 library concentration was too low for quantification.
Reliable NGS library quantification.
The DNA standards for the Illumina MiSeq platform from three different lots were used to prepare five sequential 10-fold dilutions. The diluted DNA standards were mixed with a library-specific PCR primer assay and GeneRead qPCR SYBR Green Mastermix (from three different lots) and were subjected to real-time PCR. The minimal variation in CT values for diluted DNA standards from three different lots confirms the reliability of the Gene Read Library Quant System.
High yields of library DNA with uniform coverage distribution.
[A] Genomic DNA (50 ng) was sheared using a Covaris instrument and made into an Illumina-compatible DNA library using the GeneRead Library Prep (I) Kit. Sequencing using an Illumina MiSeq instrument revealed a median coverage of 49-fold with uniform coverage distribution. [B] Genomic DNA (1 µg) from E. coli strain DH10B was sheared and used to generate an Ion Torrent-compatible DNA library using the GeneRead Library Prep (L) Kit or a kit from another supplier. After amplification for 10 cycles, both libraries were sequenced on an Ion Torrent PGM instrument and the cumulated normalized coverage was analyzed.