QIAseq Targeted RNA Extended Panels
用于基因表达图谱分析的数字RNAseq
Products
QIAseq Targeted RNA Extended Panels 旨在用于分子生物学应用。这些产品不能用于疾病诊断、预防和治疗。
特点
- 可将多达25个基因添加至预制panel中
- 每个panel最低仅需25 ng总RNA
- 在1天之内可完成“从样本制备到文库构建”的全部流程
- 采用分子条形码技术，保证了表达图谱分析的准确度
产品详情
QIAseq Targeted RNA Extended Panel是通过RNAseq进行定量基因表达图谱分析的“从样本制备到数据解读”解决方案。这些panel结合了分子条形码技术和两步法PCR的文库制备方法，可无偏差、准确地定量数字RNA测序结果。
绩效
- Accuracy: Innovative digital sequencing (incorporating molecular barcodes) eliminates PCR duplication and amplification bias to confidently detect known and novel fusions.
- Specificity: The unique combination of our proprietary primer design algorithm and rigorous testing of every primer assay guarantees high specificity and accurate results.
- Content: The QIAseq targeted RNAscan panels offer a high degree of flexibility in content and sample multiplexing. Several cataloged panels have been developed for a wide range of applications. One can also build a custom panel for specific content, or to extend the contents of an existing cataloged panel. Up to 384 samples can be multiplexed using the QIAseq indexes.
- Flexibility: Because the QIAseq targeted RNAscan panels use single primer extension, primers can be designed to detect known fusions based on characterized breakpoints or to discover novel fusions.
原理
PCR duplicates are a major issue in targeted RNA sequencing for gene fusion detection, since – through PCR amplification – they turn unique RNA molecules into identical RNA molecules that cannot be distinguished from each other. This, in turn, results in the inability to confidently detect gene fusions. To overcome the issue of PCR duplicates, the QIAseq targeted RNAscan panels use digital sequencing by incorporating molecular barcodes into the starting RNA material before any amplification takes place – thereby preserving the uniqueness of the starting RNA molecules and overcoming the issues of not only PCR duplicates and amplification bias.
程序
The entire workflow of the QIAseq Targeted RNAscan Panels – from extracted RNA to sequencing-ready libraries – can be completed in 9 hours. Extracted RNA is converted to cDNA, targets are molecularly barcoded and enriched, and libraries are constructed. Sequencing files can be fed into the QIAseq pipeline – a cloud-based data analysis pipeline – which will filter, map and align reads, as well as count unique molecular barcodes associated with targeted regions and call fusions. This data can then be fed into QCI for interpretation.
应用
- 基因表达图谱分析
- 生物标志物研究
- 全转录组测序数据验证
- 微阵列数据验证
辅助数据和图表
Workflow
Isolated RNA, as low as 20 ng, is converted to cDNA. This is followed by the library construction step, in which IL-N7 adapters, molecular barcodes, and sample indexes are incorporated into DNA fragments generated in the previous step. Library fragments now serve as templates for target enrichment using single primer extension. In this step, targets are enriched using a single gene-specific primer and a universal forward primer. The final step is library amplification and sample indexing (for dual indexing) using the IL-S5 sample index primer and a universal primer.
资源
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