QIAseq Targeted RNA Extended Panels

1. 可将多达25个基因添加至预制panel中
2. 每个panel最低仅需25 ng总RNA
3. 在1天之内可完成“从样本制备到文库构建”的全部流程
4. 采用分子条形码技术，保证了表达图谱分析的准确度

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。

1. 准确性：创新的数字测序技术（分子条形码计数法）消除了PCR复制和扩增引入的偏差，可获得准确的结果（参见图无偏差、准确的基因定量检测结果）
2. 特异性：独创性地将专有的引物设计算法和每个引物延伸实验的严格检测相结合，保证了结果的高度特异性和准确性（参见图专有的引物设计– 97%的特异性）。
3. 一致性：QIAseq Targeted RNA Panel工作流程已经过优化，可获得高度一致的测序结果，保证仪器的测序能力得到有效利用。实际上，数字测序（分子条形码）已完全消除了RNAseq计数中产生的数据波动（请参见图卓越的一致性 – 97%的实验结果均处于分子标记数中位数的20%内）。
4. 可重复性：QIASeq Targeted RNA Panel系统重复性强，在重复样本、不同批次产品和不同仪器的检测结果间存在很强的相关性，平均相关系数高于0.99，保证了检测的可靠性。
5. 灵敏性：数字RNA测序系统已经过优化，可获得极为可靠的定量检测结果，可定量25 ng总RNA之中低至100个拷贝的RNA靶标（参见图每个细胞低至0.2个拷贝数RNA的阳性测定结果）。
6. 灵活性：QIAseq panel将NGS的强大功能和qPCR的准确性合为一体。在获得高性价比数据结果的同时，可通过单次NGS多重检测多个样本。

DirectPrep 96高通量质粒DNA分离利用优化的裂解技术，无需澄清裂解液。只需通过单板以及简便的结合-洗涤-洗脱流程即可完成质粒DNA纯化。该操作方法比现有其他大多数方法更易于操作。DirectPrep 96操作流程需要在QIAvac Multiwell上完成。

• 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.

1. 基因表达图谱分析
2. 生物标志物研究
3. 全转录组测序数据验证
4. 微阵列数据验证