Qiagen/QIAseq Targeted RNA Panels/333002
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商品介绍
QIAseq Targeted RNA Panels
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Digital RNAseq for gene expression profiling
- 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 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.
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Cat No./ID:333002 QIAseq Targeted RNA Panel (12) Go to GeneGlobeKit containing wet bench-verified assays and reagents for first strand synthesis, molecular barcoding, gene-specific amplification and library preparation for targeted RNA sequencing |
Cat No./ID:333005 QIAseq Targeted RNA Panel (96) Go to GeneGlobeKit containing wet bench-verified assays and reagents for first strand synthesis, molecular barcoding, gene-specific amplification and library preparation for targeted RNA sequencing |
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
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 ofour 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 deliveraccurate 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.
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
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品牌介绍
QIAGEN是一家专业化致力于生物分子样品制备解决方案的跨国经营企业，总部位于德国。1984年，QIAGEN在德国成立，1996年在美国纽约纳斯达克上市。QIAGEN拥有超过1000项专利和认可证明，在18个国家设立了分公司，代理商服务国超过40个，在全球有超过400000的用户。QIAGEN提供的产品超过500类，包括各种试剂，耗材和自动化纯化工作站。这些产品用于样品采集，稳定，核酸或蛋白的分离，纯化和检测中，不仅广泛的应用于科研领域的各个方面，在生物技术，制药，法医研究，食品安全检测，畜牧业和分子诊断领域也得到了广泛的应用。QIAGEN产品的卓越品质和在应用中的出色表现使得其成为样品处理中标准的代名词。
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