GeneRead DNAseq NGS workflow
Targeted enrichment using multiplex PCR
Answer specific biological and clinical research questions quickly and easily with GeneRead DNAseq Panels V2, which use fast, multiplex-PCR-based targeted enrichment to allow deep sequencing of specific genes or genomic regions of interest in a wide range of asmples (see
GeneRead DNAseq Targeted Panel System). Integrated DNA quality and targeted enrichment controls, sophisticated, wet-bench verified primer design, and optimized targeted enrichment chemistry ensure high design coverage, specificity, and uniformity. More than 90% of targeted regions are covered by the panel primer design, enabling deep sequencing and the identification of low-frequency genetic variants that might otherwise go undetected.
DNA sequencing is a useful tool to detect genetic variations, including somatic mutations, SNPs, and small insertions and deletions (indels). However, sequencing the genome can be time-consuming and costly. Targeted enrichment technology enables NGS platform users to sequence specific regions of interest instead of the entire genome, thereby achieving more sensitive mutation detection and reduced costs. GeneRead DNAseq Panels V2 are designed to analyze a panel of genes related to a disease state and can be used with any major NGS platform. Targeted enrichment is particularly well suited for medium-throughput sequencers, including Life Technologies’ Ion Torrent PGM Sequencer and Illumina’s MiSeq Personal Sequencer. Wet-bench verified primer sets are available for most popular genes, as well as custom panels with any set of genes or genomic regions in the human genome.
GeneRead DNAseq Targeted Panels V2 include the most commonly mutated genes in specific human disease states (see table "GeneRead DNAseq Panels V2"). The mutations in the tumor suppressor genes and oncogenes that make up the cancer-specific and comprehensive panels are often relevant for tumor classification and warrant extensive investigation to enhance our understanding of carcinogenesis. You can also analyze a specific set of genes or genomic regions using GeneRead DNAseq Mix-n-Match or Custom Panels V2.
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Tissue-specific panels (<100 genes) |
Comprehensive panels (>100 genes) |
Clinically relevant panels |
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Breast Cancer |
Comprehensive Cancer |
Tumor Actionable Mutations |
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Colorectal Cancer |
Carrier Testing |
Clinically Relevant Tumor |
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Myeloid Neoplasms |
Cancer Predisposition |
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Liver Cancer |
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Lung Cancer |
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Ovarian Cancer |
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Prostate Cancer |
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Gastric Cancer |
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Cardiomyopathy |
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Build your own panel from QIAGEN's pool of 570 wet-bench verified genes |
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Build your own panel to target any gene or genomic region in the human genome |
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Overlapping primer sets across the exonic portions of a gene maximize target coverage and minimize non-specific amplification (see figure
PCR-enabled targeted enrichment). GeneRead DNAseq Targeted Panels V2 are designed to cover the coding sequences of any protein coding region of any gene in the human genome. The overlapping PCR products (amplicons) produced by the overlapping primer sets yield greater than 90% coverage of the targeted genes and more than 90% of sequence specificity.
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RNAseq NGS workflow
Highly efficient and selective rRNA depletion
Ribosomal RNA (rRNA) makes up 85–90% of total RNA, taking up valuable sequencing capacity and resulting in a high signal-to-noise ratio that can make detection of the RNA species of interest difficult. The GeneRead rRNA Depletion Kit effectively removes ribosomal RNA, while ensuring complete recovery of mRNA and noncoding RNA from various species, including human, mouse, and rat (see figure
GeneRead combines DNA/RNA sequence specificity and antibody binding). By improving the ratio of useful data, decreasing bias, and preserving non-coding RNA species, the kit provides high-quality RNA that is especially suited for NGS applications.
Efficient removal of all rRNA species
The GeneRead rRNA Depletion Kit efficiently removes all types of rRNA, including the 4 main ribosomal RNAs, as well as mitochondrial rRNA. QIAxcel Advanced data showing rat spleen RNA demonstrates complete removal of the 18S and 28S ribosomal RNA peaks compared to total RNA (see figure
Highly efficient rRNA removal). In this experiment, more than 99.9% of all types of ribosomal RNA were removed with the kit.
Highly specific rRNA depletion with no artificial depletion of protein-coding genes
The biotype distribution of sequenced RNA following rRNA depletion demonstrates that the GeneRead rRNA Depletion Kit effectively eliminates rRNA, while preserving more miRNA and other noncoding RNA than rRNA depletion kits from other suppliers (see figure
More effective and specific rRNA depletion). The biotype distribution of RNA prepared using kits from Suppliers E and I revealed significantly less effective rRNA elimination, while the kit from Supplier E enriched for scRNA (e.g., 7SL and Alu RNA).
With the GeneRead rRNA Depletion Kit, the distribution of multiple RNA types, such as miRNA and other noncoding RNAs, is also preserved compared to poly A
+ purification, which enriches for protein-coding RNAs. Kits from other suppliers may skew the representation of protein-coding (poly A
+) RNAs in a sample, particularly by nonspecific removal of non-ribosomal RNA. In contrast, the GeneRead rRNA Depletion Kit demonstrates greater concordance with poly purification and preserves the natural representation of other RNA species and protein-coding genes (see figures
The GeneRead rRNA Depletion Kit ensures greater conservation of the mRNA profile and
Better representation of protein coding genes).
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Automate your rRNA depletion with the QIAcube
The majority of the GeneRead rRNA Depletion Kit procedure, from hybridization and capture through to RNA cleanup, is automatable on the
QIAcube. The innovative QIAcube uses advanced technology to process QIAGEN spin columns, enabling seamless integration of automated, low-throughput sample prep into your laboratory workflow. No change of purification chemistry is required, assuring fast startup and immediate results.