Apostle MiniEnrich 羧基磁珠 (片段筛选和纯化)

Apostle’s MiniEnrich 技术已经在英国皇家化学会会刊  Analyst 发表，并被选为封面文章。

• High-resolution DNA size enrichment using a magnetic nano-platform and application in non-invasive prenatal testing.  Analyst. July 2020, 145, 5733-5739  (PDF)
  

摘要

Precise DNA sizing can boost sequencing efficiency, reduce cost, improve data quality, and even allow sequencing of low-input samples, while current pervasive DNA sizing approaches are incapable of differentiating DNA fragments under 200 bp with high resolution (<20 bp). In non-invasive prenatal testing (NIPT), the size distribution of cell-free fetal DNA in maternal plasma (main peak at 143 bp) is significantly different from that of maternal cell-free DNA (main peak at 166 bp). The current pervasive workflow of NIPT and DNA sizing is unable to take advantage of this 20 bp difference, resulting in sample rejection, test inaccuracy, and restricted clinical utility. Here we report a simple, automatable, high-resolution DNA size enrichment workflow, named MiniEnrich, on a magnetic nano-platform to exploit this 20 bp size difference and to enrich fetal DNA fragments from maternal blood. Two types of magnetic nanoparticles were developed, with one able to filter high-molecular-weight DNA with high resolution and the other able to recover the remaining DNA fragments under the size threshold of interest with >95% yield. Using this method, the average fetal fraction was increased from 13% to 20% after the enrichment, as measured by plasma DNA sequencing. This approach provides a new tool for high-resolution DNA size enrichment under 200 bp, which may improve NIPT accuracy by rescuing rejected non-reportable clinical samples, and enable NIPT earlier in pregnancy. It also has the potential to improve non-invasive screening for fetal monogenic disorders, differentiate tumor-related DNA in liquid biopsy and find more applications in autoimmune disease diagnosis.

• Apostle MiniEnrich 片段筛选磁珠可从不同长度的核酸混合物中筛选出特定目标长度的片段核酸，可广泛应用于NGS和分子生物学领域。
• postle MiniEnrich 纯化磁珠可从各种酶反应体系中高效纯化获得高质量的DNA产物， 广泛应用于NGS和分子生物学领域的工作流程，包括文库制备、PCR扩增、克隆等。 
  

功能特点:

• 新型纳米磁珠结构和具有专利的表面化学修饰
• 更有针对性地去除非目标片段
• 更强的磁响应力带来更短的操作时间和更少的磁珠残留
• 兼容现有的NGS和分子生物学操作流程

The Apostle MiniEnrich 片段筛选磁珠可从不同长度的核酸混合物中筛选出特定目标长度的片段核酸，可广泛应用于NGS和分子生物学领域。 Apostle MiniEnrich 片段筛选磁珠可用于从已打断的DNA片段、连接上测序接头的文库或经过PCR扩增的文库中选取特定目标长度的片段。 基于Apostle MiniEnrich 特有的磁珠技术，该试剂盒可以自定义筛选长度区间，有针对性地去除非目标片段，同时高效回收目标片段。

Apostle MiniEnrich 纯化磁珠可从各种酶反应体系中高效纯化获得高质量的DNA产物， 广泛应用于NGS和分子生物学领域的工作流程，包括文库制备、PCR扩增、克隆等。 基于Apostle MiniEnrich 特有的磁珠技术，相比于市面上其他产品， 该试剂盒可以更高效地去除污染物，更好地回收目标片段；该产品还可以搭配 Apostle MagTouch 2000自动化平台进行高通量自动化的核酸纯化。

对含有双链DNA梯度标准品（20-1000bp）的TE溶液进行纯化的模拟实验可见: 对比某国际品牌（红色）， Apostle MiniEnrich  纯化磁珠(蓝色)在高效回收大于150bp片段的同时， 含有更少的小片段残留。这一实验展示了Apostle MiniEnrich 纯化磁珠可以用于去除PCR反应中的引物（20-30bp）， 引物二聚体 (40-60bp)，建库过程中的接头 (50-60bp)，接头二聚体 (100-120bp)等，同时有效保留建库或其他分子生物学中的目标产物。

High-resolution DNA size enrichment using a magnetic nano-platform and application in non-invasive prenatal testing.  

Zhang et al. Analyst.  July 2020, 145, 5733-5739  (PDF)

Precise DNA sizing can boost sequencing efficiency, reduce cost, improve data quality, and even allow sequencing of low-input samples, while current pervasive DNA sizing approaches are incapable of differentiating DNA fragments under 200 bp with high resolution (<20 bp). In non-invasive prenatal testing (NIPT), the size distribution of cell-free fetal DNA in maternal plasma (main peak at 143 bp) is significantly different from that of maternal cell-free DNA (main peak at 166 bp). The current pervasive workflow of NIPT and DNA sizing is unable to take advantage of this 20 bp difference, resulting in sample rejection, test inaccuracy, and restricted clinical utility. Here we report a simple, automatable, high-resolution DNA size enrichment workflow, named MiniEnrich, on a magnetic nano-platform to exploit this 20 bp size difference and to enrich fetal DNA fragments from maternal blood. Two types of magnetic nanoparticles were developed, with one able to filter high-molecular-weight DNA with high resolution and the other able to recover the remaining DNA fragments under the size threshold of interest with >95% yield. Using this method, the average fetal fraction was increased from 13% to 20% after the enrichment, as measured by plasma DNA sequencing. This approach provides a new tool for high-resolution DNA size enrichment under 200 bp, which may improve NIPT accuracy by rescuing rejected non-reportable clinical samples, and enable NIPT earlier in pregnancy. It also has the potential to improve non-invasive screening for fetal monogenic disorders, differentiate tumor-related DNA in liquid biopsy and find more applications in autoimmune disease diagnosis.