QIAseq Targeted RNA Custom Panels and Indices

For highly accurate gene expression quantification using UMIs

S_1084_5_GEN_V2
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QIAseq Targeted RNA Custom Panel (12)

Cat. No. / ID:  333022

Kit containing reagents for first strand synthesis, molecular tagging, gene-specific amplification and library preparation for targeted RNA sequencing; custom panel for 12 samples
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PanelIndex
QIAseq Targeted RNA Custom Panel
QIAseq Targeted RNA 12-Index
QIAseq Targeted RNA 96-Index
Samples
12
96
384
QIAseq Targeted RNA Custom Panels and Indices are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
Configure at GeneGlobe
Find or custom design the right target-specific assays and panels to research your biological targets of interest.

Features

  • Choose from comprehensive panels or select up to 1000 genes and build a custom panel
  • UMIs (unique molecular indices) ensure accurate gene expression profiling
  • Suitable as an alternative to qPCR for gene expression from as little as 25 ng of total RNA
  • Includes access to GeneGlobe data analysis portal for read alignment, data normalization and gene expression analysis
  • Available for Illumina and Thermo-Fisher NGS instruments

Product Details

QIAseq Targeted RNA Panels have been developed as a Sample to Insight solution for quantitative gene expression profiling using RNA-seq. These panels integrate unique molecular indices (UMIs) and a two-stage PCR-based library preparation to deliver unbiased and accurate quantification from human, mouse and rat samples.

QIAseq Targeted RNA Indices are for indexing samples for targeted RNA sequencing and primers necessary for sequencing RNA libraries generated by QIAseq Targeted RNA Panels.

Performance

See figures

Principle

Traditional RNA sequencing methods suffer from PCR duplication and amplification bias, resulting in inaccurate gene expression analysis. By introducing UMIs (unique molecular indices) 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 Panel workflow begins with converting total RNA into cDNA (see figure  "Simple procedure"). The workflow requires minimal RNA input: typically 25 ng of total RNA can be used. No enrichment or rRNA depletion steps are necessary. The molecular indexing step makes use of a gene-specific primer (GSP1) which contains a 12-base UMI in a multiplex primer panel (targeting 12–1000 genes). After the molecular indexing step, the uniquely tagged cDNA is bead purified 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.

Purchase of QIAseq targeted RNA Panels provides access to data analysis tools at QIAGEN’s GeneGlobe portal. These include read alignment and data normalization, differential gene expression at no additional costs.

See figures

Applications

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

Supporting data and figures

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