QIAseq Targeted RNA Extended Panels

Digital RNAseq for gene expression profiling

Features

  • 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

Products

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.
Image
QIAseq Targeted RNA Extended Panel (12)

Cat. No. / ID: 333012

Kit containing reagents for first strand synthesis, molecular tagging, gene-specific amplification and library preparation for targeted RNA sequencing; extended panel for 12 samples
Image
QIAseq Targeted RNA Extended Panel (96)

Cat. No. / ID: 333015

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

Product Details

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. 


Need a quote for your research project or would you like to discuss your project with our specialist team? Contact Us

 

Performance


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

Resources

Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.
Kit Handbooks (1)

FAQ

Are there important considerations for plasma generation and urine handling?

It is strongly advised to follow the recommendations for preparing sample material provided in the corresponding Protocol Sheet to ensure reliable results.

Plasma: It is recommended to perform plasma separation immediately after blood collection when using EDTA or citrate as anticoagulant to prevent the release of genomic DNA into the plasma fraction.

Urine: Because circulating cell-free DNA in non-stabilized urine samples is rapidly degraded after sample collection due to high nuclease activity, eluates may contain no DNA or exhibit low DNA concentration. Therefore, it is recommended to stabilize urine samples. Even when using stabilized urine, it is recommended to perform a centrifugation step immediately after stabilization to prevent the release of genomic DNA from cells. Alternatively, non-stabilized urine samples can be processed immediately after collection and centrifugation using ATL-pretreatment and automated DNA extraction as described in the corresponding Protocol Sheet.

FAQ ID - 3699