TARGET-Seq: A Protocol for High-Sensitivity Single-Cell Mutational Analysis and Parallel RNA Sequencing.
TARGET-Seq: A Protocol for High-Sensitivity Single-Cell Mutational Analysis and Parallel RNA Sequencing.
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DOI：
10.1016/j.xpro.2020.100125
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发表时间：
2020-12-18
期刊：
影响因子：
--
通讯作者：
Mead AJ
中科院分区：
文献类型：
--
作者：
Rodriguez-Meira A;O'Sullivan J;Rahman H;Mead AJ
单细胞RNA测序技术是解决肿瘤内异质性的理想选择。然而，关键突变热点的覆盖率的缺乏已经排除了来自相同单细胞的遗传和转录读数的相关性。为了克服这一点，我们开发了TARGET-seq，这是一种用于TARGETed高灵敏度单细胞突变分析的方案，具有极低的等位基因脱落率，并行RNA测序和细胞表面蛋白质组学。在这里，我们为TARGET-seq提供了一个详细的分步协议，包括故障排除提示，自动化方法以及库的高通量多路复用方法。有关本方案使用和执行的完整详细信息，请参阅。TARGET-seq提供高灵敏度的单细胞突变分析和RNA测序TARGET-seq解决了遗传和分子肿瘤内异质性机器人平台中的全自动方案用于高通量分析单细胞RNA测序技术是解决肿瘤内异质性的理想选择。然而，关键突变热点的覆盖率的缺乏已经排除了来自相同单细胞的遗传和转录读数的相关性。为了克服这一点，我们开发了TARGET-seq，这是一种用于TARGETed高灵敏度单细胞突变分析的方案，具有极低的等位基因脱落率，并行RNA测序和细胞表面蛋白质组学。在这里，我们为TARGET-seq提供了一个详细的分步协议，包括故障排除提示，自动化方法以及库的高通量多路复用方法。
Single-cell RNA-sequencing technologies are ideally placed to resolve intratumoral heterogeneity. However, the lack of coverage across key mutation hotspots has precluded the correlation of genetic and transcriptional readouts from the same single cell. To overcome this, we developed TARGET-seq, a protocol for TARGETed high-sensitivity single-cell mutational analysis with extremely low allelic dropout rates, parallel RNA SEQuencing, and cell-surface proteomics. Here, we present a detailed step-by-step protocol for TARGET-seq, including troubleshooting tips, approaches for automation, and methods for high-throughput multiplexing of libraries. For complete details on the use and execution of this protocol, please refer to. TARGET-seq provides high-sensitivity single-cell mutational analysis and RNA-sequencing TARGET-seq resolves genetic and molecular intratumoral heterogeneity Fully automated protocol in robotic platforms for high-throughput analysis Single-cell RNA-sequencing technologies are ideally placed to resolve intratumoral heterogeneity. However, the lack of coverage across key mutation hotspots has precluded the correlation of genetic and transcriptional readouts from the same single cell. To overcome this, we developed TARGET-seq, a protocol for TARGETed high-sensitivity single-cell mutational analysis with extremely low allelic dropout rates, parallel RNA SEQuencing, and cell-surface proteomics. Here, we present a detailed step-by-step protocol for TARGET-seq, including troubleshooting tips, approaches for automation, and methods for high-throughput multiplexing of libraries.
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影响因子：
48
作者：
Picelli, Simone;Bjorklund, Asa K.;Sandberg, Rickard
通讯作者：
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影响因子：
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作者：
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DOI：
10.1093/bioinformatics/btp352
发表时间：
2009-08-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
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作者：
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1000 Genome Project Data Processing Subgroup
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作者：
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