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PyroMark CpG Assay (200)
PCR and sequencing primers for Pyrosequencing analysis of gene-specific CpG methylation after DNA bisulfite conversion (200 reactions; tube format)
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Varies
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PyroMark CpG Assay 96 well (50)
PCR and sequencing primers for Pyrosequencing analysis of gene-specific CpG methylation after DNA bisulfite conversion (50 reactions; plate format); minimum order of 24 assays per plate
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Varies
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PyroMark CpG Assay 96 well (200)
PCR and sequencing primers for Pyrosequencing analysis of gene-specific CpG methylation after DNA bisulfite conversion (200 reactions; plate format); minimum order of 24 assays per plate
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Varies
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PyroMark CpG Assays are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
Analysis of ESR1.|High PCR success rate, even with bisulfite converted DNA.|High Pyrosequencing success rate of predesigned PyroMark CpG Assays.|Ready-to-use methylation assays.|
[A] The GeneGlobe interface for PyroMark CpG Assays affords access to multiple targets in CpG islands, which allows the user to pick and choose the particular regions in a CpG island to analyze. [B] Pyrosequencing data show the analysis of two different positions within the CpG island associated with the gene ESR1.|The PCR performance of PyroMark CpG Assays for 310 randomly selected genes was evaluated. A total of 305 assays (98%) resulted in the predicted PCR amplicon, usable as template for subsequent Pyrosequencing.|The Pyrosequencing performance of PyroMark CpG Assays for 310 randomly selected genes was evaluated. A total of 305 assays (98%) resulted in the predicted PCR amplicon, usable as template for subsequent Pyrosequencing. From these assays, 281 resulted in successful Pyrosequencing results. Of these successful assays, 229 (75.1%) required no further review by the user, and 52 (17%) were successful after user review. Only 24 (7.9%) evaluated assays failed.|The GeneGlobe interface for searching and ordering PyroMark CpG Assays provides access to multiple targets in CpG islands, allowing users to pick and choose the exact regions in a CpG island for analysis. By combining different assays, users can quantify methylation of as many target sites as needed, whether these are consecutive or not.|
Principle
PyroMark CpG Assays可对基因组的特异性靶基因进行甲基化分析。这些预制的甲基化分析试剂使用先进的运算法则设计，确保对亚硫酸氢盐转化后DNA最优化的分析。使用PyroMark CpG Assay节约了分析设计的时间，并最大程度确保实验成功。PyroMark CpG Assays与PyroMark产品线的其他产品，如PCR产物和测序试剂配合使用，可提高焦磷酸测序工作流程的效率，实现对人类和小鼠基因组所有CpG位点的甲基化分析。
优化的引物设计
PyroMark CpG Assays中提供的PCR和测序引物都使用先进的、功能强大的运算法则设计。所得引物可针对人和小鼠全基因组进行检测，引物浓度为最佳浓度，提高PCR的特异性。这些试剂配合PyroMark PCR Kit的专用试剂使用，可优化结果，提高测序模板的产量和品质，提高分析的可靠性。
GeneGlobe的优势
QIAGEN的GeneGlobe Web portal是一站式在线数据库，可按照不同的基因或信号通路，选择我们多种多样的分析方法和技术。GeneGlobe具有最先进的搜索和设计技术、最新的序列信息和创新的技术。只需输入您感兴趣的基因或信号通路的名称，GeneGlobe将会为您展示在表观遗传学或其他您感兴趣的领域如何优化您的研究。
Procedure
在焦磷酸测序之前，从亚硫酸氢盐转化DNA获得荧光素标记的PCR产物。荧光素标记的PCR产物结合到覆盖有链酶亲和素的Sepharose珠上。在PyroMark Vacuum Workstation上，这些珠子吸附到真空容器中，充分洗涤，产生适合测序的单链DNA。PyroMark CpG Assays提供预制的DNA甲基化分析试剂，用于焦磷酸测序。只需在GeneGlobe portal中点击几下即可获得超过60,000个人和小鼠全基因组分析。每个分析即含有产生荧光素标记的PCR产物所需的引物，也含有焦磷酸测序反应所需的引物。
Applications
PyroMark CpG Assays能够高度准确的对小鼠和人类CpG位点进行定量甲基化分析，适合于表观遗传学研究。