QIAseq Targeted RNA Panels

Digital RNAseq for gene expression profiling

Features

  • Molecular barcodes remove PCR and library construction bias
  • Start with only 25 ng of total RNA
  • Simple 1 day library construction workflow
  • Use with any illumina or Thermo-Fisher NGS instrument
QIAseq Targeted RNA Panel (12)

Cat. No. / ID: 333002

Kit containing wet bench-verified assays and reagents for first strand synthesis, molecular barcoding, gene-specific amplification and library preparation for targeted RNA sequencing
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samples
12
96
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QIAseq Targeted RNA Panels are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

QIAseq Targeted RNA 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. QIAseq Targeted Panels are wet-bench verified to ensure the highest quality of results.


Performance


See figures

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 housekeeping gene (HKG) assays are used to normalize data, thereby making sample-to-sample and run-to-run comparisons possible.

 

See figures

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.
See figures

Applications

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

Supporting data and figures

Resources

Scientific Posters (1)
Poster for download
Brochures & Guides (3)
State-of-the-art technologies to fast-track and streamline NGS workflows
Kit Handbooks (1)

Publications

Assessment of mitochondrial function following short- and long-term exposure of human bronchial epithelial cells to total particulate matter from a candidate modified-risk tobacco product and reference cigarettes.
Malinska D; Szymański J; Patalas-Krawczyk P; Michalska B; Wojtala A; Prill M; Partyka M; Drabik K; Walczak J; Sewer A; Johne S; Luettich K; Peitsch MC; Hoeng J; Duszyński J; Szczepanowska J; van der Toorn M; Wieckowski MR;
Food Chem Toxicol; 2018; 115 :1-12 2018 Feb 13 PMID:29448087
A function-blocking CD47 antibody modulates extracellular vesicle-mediated intercellular signaling between breast carcinoma cells and endothelial cells.
Kaur S; Elkahloun AG; Singh SP; Arakelyan A; Roberts DD;
J Cell Commun Signal; 2017; 12 (1):157-170 2017 Nov 29 PMID:29188480
An Atlas of Human Regulatory T Helper-like Cells Reveals Features of Th2-like Tregs that Support a Tumorigenic Environment.
Halim L; Romano M; McGregor R; Correa I; Pavlidis P; Grageda N; Hoong SJ; Yuksel M; Jassem W; Hannen RF; Ong M; Mckinney O; Hayee B; Karagiannis SN; Powell N; Lechler RI; Nova-Lamperti E; Lombardi G;
Cell Rep; 2017; 20 (3):757-770 2017 Jul 18 PMID:28723576
Chemoprevention with Cyclooxygenase and Epidermal Growth Factor Receptor Inhibitors in Familial Adenomatous Polyposis Patients: mRNA Signatures of Duodenal Neoplasia.
Delker DA; Wood AC; Snow AK; Samadder NJ; Samowitz WS; Affolter KE; Boucher KM; Pappas LM; Stijleman IJ; Kanth P; Byrne KR; Burt RW; Bernard PS; Neklason DW;
Cancer Prev Res (Phila); 2017; 11 (1):4-15 2017 Nov 6 PMID:29109117
A Role for Iodide and Thyroglobulin in Modulating the Function of Human Immune Cells.
Bilal MY; Dambaeva S; Kwak-Kim J; Gilman-Sachs A; Beaman KD;
Front Immunol; 2017; 8 :1573 2017 Nov 15 PMID:29187856