Ripple Biosolutions, Seattle, Washington, United States of America.
PlasmaLab International, Everett, Washington, United States of America.
PLoS One. 2020 Jun 1;15(6):e0231854. doi: 10.1371/journal.pone.0231854. eCollection 2020.
UltraPrep is an open-source, two-step method for purification of cell-free DNA that entails extraction of total DNA followed by size-selective enrichment of the smaller fragments that are characteristic of DNA originating from fragmentation between nucleosome. The advantages of the two related protocols that are described are that they can easily accommodate a wide range of sample input volumes, they rely on simple, magnetic bead-based technology, the yields of cfDNA are directly comparable to the most popular methods for cfDNA purification, and they dramatically reduce the cost of cfDNA isolation relative to currently available commercial methods. We provide a framework for physical and molecular quality analysis of purified cfDNA and demonstrate that the cfDNA generated by UltraPrep meets or exceeds the quality metrics of the most commonly used procedure. In addition, our method removes high molecular weight genomic DNA (hmwgDNA) that can interfere with downstream assay results, thereby addressing one of the primary concerns for preanalytical collection of blood samples.
UltraPrep 是一种开源的两步法，用于纯化无细胞 DNA，包括提取总 DNA，然后进行大小选择，富集小片段，这些小片段是核小体之间断裂产生的 DNA 的特征。所描述的两种相关方案的优势在于，它们可以轻松适应广泛的样品输入体积，它们依赖于简单的基于磁珠的技术，cfDNA 的产量与最受欢迎的 cfDNA 纯化方法直接可比，并且与目前可用的商业方法相比，cfDNA 分离的成本大大降低。我们提供了一个用于纯化 cfDNA 的物理和分子质量分析框架，并证明 UltraPrep 产生的 cfDNA 满足或超过最常用方法的质量指标。此外，我们的方法去除了可能干扰下游检测结果的高分子量基因组 DNA（hmwgDNA），从而解决了血液样本预分析采集的主要问题之一。
