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多重检测PCR扩增子

 

  • 经过长达10 kb扩增子的验证
  • 通过格式化允许2到96个样本的多重扩增
  • 根据经过验证或可更换的引物设计,采用不同的流程
  • 模板制备的成本降低90倍
  • 模板制备的时间降低50%
  • 设置后适用于自动化液体处理装置

特定区域的直接PCR扩增是一种强大的技术,适用于靶向测序。当与多重检测能力相结合时,PCR扩增就成为一种研究多个样本的高效方法。PacBio为多重扩增提供了两种添加条形码的策略。第一种方法是在PCR过程中向扩增子的末端添加条形码,随后将标准的SMRTbell接头与带有条形码的扩增子连接。整个流程如下图所示。

barcoded-universal-primer-workflow

第二种方法是在SMRTbell接头上添加条形码,然后将扩增子与带有条形码的SMRTbell接头连接。第一种方法更为经济高效,因为条形码是添加到扩增子上,并可在SMRTbell模板制备步骤之前混合样本。条形码标记的接头流程如下图所示。

barcoded-adapter-workflow
对于经过验证的PCR系统,建议使用第二种方法,因为扩增子是利用未经修饰的引物产生的。这两种方法的流程和好处在这个条形码方案的产品指南中有讨论。

若有意了解如何运用我们的SMRT消耗品来支持您的研究,请联系我们