BD Rhapsody Onco-BC 乳癌靶向检测panel

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BD Rhapsody Onco-BC 乳癌靶向检测panel：
经验证的基因panel专注于人类乳腺癌生物学
有针对性的方法提高了实验效率
可重复的检测方法可用于各种实验

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BD Rhapsody Onco-BC 乳癌靶向检测试剂盒（panel）

英文名称：BD Rhapsody™  Onco-BC Targeted Panel

中文名称：BD Rhapsody Onco-BC 乳癌靶向检测试剂盒（panel）

用途：与BD Rhapsody单细胞表达分析仪配合使用，检测与乳腺癌和免疫细胞相关的 400 个基因

乳腺癌生物学因复杂的基因表达，细胞异质性和患者间的差异而变得复杂。为了解决这一复杂性，BD Rhapsody™OncoBC靶向检测panel使用BD Rhapsody单细胞分析系统为单细胞乳腺癌研究提供了一种简化的检测方法。 通过集中分析与乳腺癌生物学zui相关的基因和标志物，减少测序通量，并且降低了与整个转录组测序相关时间和测序成本。

BD Rhapsody™  Onco-BC Targeted Panel特点

经验证的基因panel专注于人类乳腺癌生物学

有针对性的方法提高了实验效率

可重复的检测方法可用于各种实验

欢迎您咨询华雅再生医学旗舰公司：红荣微再（上海）生物工程技术有限公司  ：1500 1904 520。红荣微再-客服:  经销商专员

BD Rhapsody单细胞分析系统

BD Rhapsody单细胞分析系统是一种新型的单细胞mRNA表达分析仪器。它能捕获并分离细胞，通过磁珠对细胞和mRNA转录本进行分类扩增以供测序。作为单细胞RNA-seq的新方法，与传统的全转录组扩增（WTA）相比，BD Rhapsody单细胞分析系统可以同时处理12个样本，能够对成千上万个单细胞的数百个表达基因进行数字定量，极大程度节省实验时间和降低测序成本。

BD Rhapsody单细胞分析系统可以定制靶向组合（panel），靶向方法有助于提高单细胞测序效率，并推动对当今一些挑战性的科学领域的进一步探索。BD经验证的检测试剂盒（panel）显示稳定且可重现的细胞异质性阵列，可用于探索与众多应用相关的细胞类型。BD Rhapsody 系统包括以下检测试剂盒（panel）：

□ BD Rhapsody Onco-BC 靶向检测试剂盒（panel） – 与乳腺癌和免疫细胞相关的 400 个基因

□ BD Rhapsody 免疫反应检测试剂盒（panel） – 免疫细胞类型谱分析

□ BD Rhapsody  T-细胞靶向检测试剂盒（panel）

□ BD Rhapsody 补充检测试剂盒（panel）

□ BD Rhapsody 自定义检测试剂盒（panel）

BD Rhapsody Onco-BC 靶向检测试剂盒（panel）

Panel designed by scientists, for scientists

The customizable and validated BD Rhapsody Onco-BC Targeted Panel was created from collaborations with clinical oncologists.

The panel content combines data from experimental and database sources, chosen to include genes with known involvement in cancer progression, proliferation, epithelial-tomesenchymal transition (EMT) and cancer stem cells. Including PAM50 genes aids molecular subtyping of breast cancer cells¹,while immunophenotyping and stromal cell markers enable comprehensive surveys of the complex tumor microenvironment.

科学家为科学家设计的Panel

可定制且经过验证的BD Rhapsody Onco-BC乳腺癌靶向Panel是由临床肿瘤学家合作开发的。

BD Rhapsody Onco-BC乳腺癌靶向Panel结合了来自实验和数据库来源的数据，选择的数据包括已知参与癌症progression，扩增，上皮 - 间质转化（EMT）和癌症干细胞的基因。 包括PAM50基因有助于乳腺癌细胞的分子亚型¹，而免疫分型和基质细胞标志物可以对复杂的肿瘤微环境进行全面调查。

Tailored libraries for targeted sequencing

The BD Rhapsody Onco-BC Targeted Panel is designed for the BD Rhapsody system, and uses multiplex PCR for detecting up to 389 genes. After cells are lysed in the BD Rhapsody Cartridge,

beads hybridized with mRNA are magnetically retrieved for cDNA synthesis. Primers included in the BD Rhapsody Onco-BC Targeted Panel and BD Rhapsody Supplemental Panel are used for several rounds of gene-specific, nested PCR for library construction. Final PCR products for sequencing contain adapters, a cell label, a unique molecular index (UMI), and up to 400 bp of the 3’ end of a target gene. Assay products may be sequenced on Illumina® MiniSeq, MiSeq, NextSeq, HiSeq2500 and HiSeq4000 sequencers.

靶向测序的定制文库

BD Rhapsody Onco-BC乳癌靶向Panel专为BD Rhapsody系统设计，使用多重PCR检测多达389个基因。细胞在BD Rhapsody Cartridge中裂解后，与mRNA杂交的磁珠被磁性回收用于cDNA合成。包含在BD Rhapsody Onco-BC靶向Panel和BD Rhapsody Supplemental Panel 中的引物用于多轮基因特异性巢式PCR，用于构建文库。用于测序的zui终PCR产物含有接头，细胞标记（label），*的分子索引（unique molecular index，UMI）和3'末端高达400bp的靶基因。检测产物可以在Illumina®MiniSeq，MiSeq，NextSeq，HiSeq2500和HiSeq4000测序仪上进行测序。

Figure 1. cDNA synthesis and library construction chemistry with the BD Rhapsody Onco-BC Targeted Panel.

图1.BD Rhapsody Onco-BC Targeted Panel的cDNA合成和文库构建原理

Your target genes, delivered your way

Researchers may select up to 100 additional genes for analysis,with the BD Rhapsody Supplemental Panel. Using BD’s primer design software, primers for supplemental gene targets are designed to be compatible with pre-designed base panels.

Supplemental primers are shipped to you and combined with the BD Rhapsody Onco-BC Targeted Panel for library preparation.

专属目标基因定制

研究人员可以经由BD Rhapsody Supplemental Panel选择zui多100个额外基因分析。使用BD的引物设计软件，补充目标基因的引物可以与预设基础panel兼容。

Remarkable panel performance—single-cell suspensions from a metastatic breast cancer sample

In the following example, cryopreserved single-cell suspensions from a disaggregated metastatic breast cancer patient sample were sorted using the BD FACSAria™ system to enrich for live cells. 4,409 cells derived from a lymph node metastasis were profiled using the BD Rhapsody system. Fifteen cell types were detected, including four subpopulations of breast cancer cells and a diverse tumor microenvironment of stromal and major immune cell types. Because the analysis focused on just five percent of the transcriptome, the amount of sequencing is drastically lowered.

的panel性能—转移性乳腺癌样本的单细胞悬液

在下面的实例中，使用BD FACSAria ^TM系统分选来自disaggregated转移性乳腺癌患者样品的冻存单细胞悬浮液以富集活细胞。使用BD Rhapsody系统对来源于淋巴结转移的4,409个细胞进行分析。检测到15种细胞类型，包括4种乳腺癌细胞亚群和多种肿瘤微环境（间质细胞和主要免疫细胞组成）。由于分析只集中在转录组的5％，因此测序通量极大降低。

Table 1: Example genes by category

Pathway # of genes in panel

Cell adhesion 23；Cell cycle 44；Chemokine/chemokine receptor,cytokine/cytokine receptor 53；

Signal transduction 56；Oncogenes, tumor suppressors 10；Tumor immunity, immune checkpoint,

immune cell activation 24；Surface antigen 32；Other* 147（ Angiogenesis; cell death; complement system; structural; transport; DNA repair;epithelial-to-mesenchymal transition; hormone receptor; immunoglobulin; lncRNA;metabolism; multi-drug resistance; protease; transcription factor）

表1 参考基因

其他：血管生成; 细胞死亡; 补充体系;结构; 运输; DNA修复;上皮间质转化; 激素受体;免疫球蛋白;lncRNA;代谢; 多重耐药性;蛋白酶; 转录因子

表2：BD Rhapsody Onco-BC Targeted Panel metrics

BD Rhapsody Onco-BC乳癌靶向检测panel应用实例

转移性乳腺癌样本分析

Analysis of a metastatic breast cancer sample

Figure 2. Example data from analysis of a lymph node containing metastatic BrCa cells using the BD Rhapsody Onco-BC Targeted Panel. (a) tSNE projection of 4,409 cells annotated by cell type and (b) box plots of marker gene expression for each major cell type show complex tumor heterogeneity.

图2.使用BD Rhapsody Onco-BC靶向Panel分析含有转移性BrCa细胞淋巴结的实例数据。 （a）细胞类型注释的4,409个细胞的tSNE projection和（b）每种主要细胞类型的标志基因box plots显示复杂的肿瘤异质性。

Learn more about the power of targeted single-cell analysis

The BD Rhapsody Onco-BC Targeted Panel brings single-cell analysis within reach of all breast cancer researchers, by combining increased sequencing efficiency with a flexible approach to your projects. Contact us to find out how the BD Rhapsody panel can help you further the understanding of breast cancer biology.

了解更多单细胞靶向分析功能

BD Rhapsody Onco-BC Targeted Panel通过将更高的测序效率与灵活的方法结合到您的项目中，使所有乳腺癌研究人员都能够进行单细胞分析。，了解BD Rhapsodypanel如何帮助您进一步了解乳腺癌生物学。

参考文献

1. Parker JS, Mullins M, Cheang MCU, et al. Supervised Risk Predictor of Breast Cancer Based on Intrinsic Subtypes. Journal of Clinical Oncology. 2009;27(8):1160-1167.

欢迎您咨询华雅再生医学旗舰公司：红荣微再（上海）生物工程技术有限公司  ：1500 1904 520。红荣微再-客服:  经销商专员