用多重测序降低你的每份样本的成本
我们的多路复用选项允许大量的样品文库在SMRT细胞上同时被汇集和测序。在针对特定的基因组区域或处理较小的基因组时，样品多路复用很有用。
通过在双链DNA的两端加入PacBio特定的条形码，对样品进行编码，以便在数据分析前对读数进行识别和分类。条形码位于发夹适配器和样品插入物之间。
多重化支持三个选项，在目标扩增或文库制备过程中灵活地加入独特的样品标识符，称为条形码。这是通过单步PCR的条码引物、通过两步PCR的标签条码引物和条码悬臂适配器实现的。
在(1)PCR过程中，通过包含条形码的目标特定引物或(2)使用条形码通用引物的两步PCR将条形码引物添加到扩增子中。
通过(3)将条形码SMRTbell适配器连接到模板上，将条形码添加到样品中。
在Sequel、Sequel II和IIe系统上设置复用功能
在PacBio Sequel系统上进行复用的条形码策略取决于应用。所有的工作流程都可以包括条形码，以唯一地识别样品，但在如何将条形码纳入SMRTbell模板上有所不同。下面按应用列出了推荐的条形码、程序、订购单和解复用所需的FASTA文件的清单。
全基因组测序
随着PacBio Sequel系统吞吐量的增加，全基因组测序的复用能力也相应增长，特别是在微生物测序、从头组装和结构变化方面。了解更多关于全基因组测序应用的信息，请看下面关于复用的细节。
Sequel II and IIe systems
Sequel system
RNA sequencing
The Iso-Seq method is the PacBio solution for sequencing full-length transcripts,the Iso-Seq express workflow is a one-day protocol that requires only 60–300 ng of total RNA and supports multiplexing of different tissues. Learn more about the RNA sequencing application and see details on multiplexing below:
Sequel II and IIe systems
Sequel system
Metagenomics
Multiplex options are available on the PacBio Sequel systems for both full-length 16S sequencing and shotgun metagenomics profiling applications. Learn more about our metagenomic applications for characterizing microbial communities and see details on multiplexing below:
Sequel II and IIe systems
Sequel system
Targeted sequencing
Depending on the amplicon size, multiplex up to 384 barcodes for targeted sequencing. Learn more about the targeted sequencing applications and see details on multiplexing below:
Sequel II and IIe systems
Sequel system
SMRT analysis barcode analysis overview and requirements
SMRT Link includes a streamlined end-to-end demultiplex barcodes analysis. Overview and details of available features are described here.
Documentation
