QIAseq Targeted RNA Extended Panels

Digital RNAseq for gene expression profiling

Add up to 25 genes to a catalog panel
Use only 25 ng of total RNA for each panel
Go from sample to sequence-ready library in 1 day
Molecular barcodes ensure accurate expression profiling

QIAseq Targeted RNA Extended Panels have been developed as a Sample to Insight solution for quantitative gene expression profiling using RNAseq. These panels integrate molecular barcode technology and a two-stage PCR-based library preparation to deliver unbiased and accurate quantification for your digital RNA sequencing results.

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Cat No./ID: 333012

QIAseq Targeted RNA Extended Panel (12)

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Kit containing reagents for first strand synthesis, molecular tagging, gene-specific amplification and library preparation for targeted RNA sequencing; extended panel for 12 samples

Cat No./ID: 333015

QIAseq Targeted RNA Extended Panel (96)

Go to GeneGlobe

Kit containing reagents for first strand synthesis, molecular tagging, gene-specific amplification and library preparation for targeted RNA sequencing; extended panel for 96 samples

QIAseq Targeted RNA Extended Panels are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

Performance

Accuracy: Innovative digital sequencing (molecular barcode counting) eliminates PCR duplication and amplification bias to deliver the most accurate results (see figure Unbiased and accurate gene quantification) .
Specificity: The unique combination of our proprietary primer design algorithm and rigorous testing of every primer assay guarantees high specificity and accurate results (see figure Proprietary primer design delivers gene-specific amplicons – 97% specificity).
Uniformity: The QIAseq Targeted RNA Panel workflow has been optimized to deliver highly uniform sequencing results, to ensure sequencing capacity is utilized very efficiently. In fact, digital sequencing (molecular barcodes) entirely remove this variation in RNAseq counting (see figure Unmatched uniformity – 97% of assays are within 20% of median molecular tag counts).
Reproducibility: The QIASeq Targeted RNA Panel system demonstrates strong correlations across technical replicates, product lots and instruments with correlation coefficients averaging above >0.99, to ensure reliable detection of differences in expression between biological samples
Sensitivity: Digital RNA sequencing system is optimized to deliver highly reliable quantification down to ~100 copies of an RNA target in 25 ng total RNA (see figure Positive results with as little as 0.2 copies of RNA per cell).
Flexibility: QIAseq panels combine the power of NGS with the accuracy of qPCR to allow multiplexing of several samples per NGS while delivering cost-effective results (see figure Simple procedure).

Principle

Traditional RNA sequencing methods suffer from PCR duplication and amplification bias, resulting in inaccurate gene expression analysis. By introducing molecular barcodes before any amplification takes place, QIAseq Targeted RNA Panels are able to eliminate this issue to deliver accurate and digital quantification of genes (see figure Unbiased and accurate gene quantification).

A unique feature of the QIAseq Targeted RNA Panels is the set of built-in control assays. The gDNA assays control for any gDNA contamination in the RNA sample to ensure reproducible results. The housekeping gene (HKG) assays are used to normalize data, thereby making sample-to-sample and run-to-run comparisons possible.

Procedure

The QIAseq Targeted RNA Panels workflow begins with converting total RNA into cDNA (see figure Simple procedure). The workflow requires minimal RNA input: as little as 25 ng total RNA can be used. No enrichment or depletion steps are necessary. The molecular barcoding step makes use of molecularly barcoded gene-specific primer (GSP1) in a multiplex primer panel (targeting 12-1000 genes) and an input of 20ng of cDNA equivalent (cDNA made from 20 ng of total RNA). After the barcoding step, the uniquely tagged cDNA is purified over beads to remove residual primers, and a PCR is set up with a second pool of gene-specific adapter primers (GSP2) and the RS2 primer, which primes off of a common tag on the GSP1 primers. This reaction insures that intended targets are enriched sufficiently to be represented in the final library. The number of cycles is kept to a minimum to keep PCR-induced variations in amplification to a low level (any variations are easily corrected and accounted for with the molecular barcodes). Another quick cleanup with beads is performed, and a universal PCR is run with RS2 and FS2 primers, which also adds sample-indexing barcodes to each sample. A final cleanup with beads is performed and the library is complete, and ready for quantification and sequencing.
An integral component of the QIAseq Targeted RNA Panels is data analysis and insight. Data analysis modules have been developed that are comprehensive, yet easy to use. Using these modules require no bioinformatics expertise. Starting with raw reads directly off the sequencer, the QIAseq targeted RNA data analysis tools at QIAGEN’s GeneGlobe portal, provide you with gene counts and fold changes, as well as links for pathway analysis.

Applications

Gene expression profiling
Biomarker research
Confirmation of whole transcriptome sequencing data
Confirmation of microarray data

Product Resources

Kit Handbooks (1)

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	QIAseq Targeted RNA Panels Handbook	Show details

Instrument Technical Documents (2)

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	QIAseq Targeted RNA Panels Template for MiSeq	Show details
	QIAseq Targeted RNA Panels Template for NextSeq	Show details

Brochures & Guides (2)

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	Exploring new frontiers with next-generation sequencing	Show details
	NGS for liquid biopsy research	Show details

Safety Data Sheets (2)

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	MSDS QIAseq Targeted RNA Extended Panel (12)
	MSDS QIAseq Targeted RNA Extended Panel (96)