Dunker J, Larsson U, Petersson D, Forsell J, Schiller A L, Alderborn A, Berg L M
Pyrosequencing AB, Uppsala, Sweden.
Biotechniques. 2003 Apr;34(4):862-6, 868. doi: 10.2144/03344pf01.
Solid-phase techniques have facilitated the handling of biochemical analytes. This has stimulated the development of systems by which large sample panels can be analyzed with high levels of security and quality. We describe a sample transfer device based on the principle of vacuum filtration, which enables parallel handling of 96 samples of analytes bound to Sepharose beads. The tool was employed for strand separation of DNA samples, by attracting the beads to filter probes while passing them between the reagent solutions. The samples were analyzed using Pyrosequencing technology and proved to yield genotyping results of high quality. The presented sample preparation procedure provides an important link in the development of integrated systems for rapid genetic analysis at a low cost. In addition, the same filter could be reused extensively with very low risk for detectable cross-contamination between assays and without any reduction in processing capacity, thus further reducing the cost per analyzed sample.
固相技术促进了生化分析物的处理。这推动了相关系统的发展,借助这些系统可以对大量样本进行高度安全且高质量的分析。我们描述了一种基于真空过滤原理的样本转移装置,它能够并行处理96个与琼脂糖珠结合的分析物样本。该工具用于DNA样本的链分离,通过在试剂溶液之间传递珠子时将其吸引到过滤探针上实现。使用焦磷酸测序技术对样本进行分析,结果证明能产生高质量的基因分型结果。所介绍的样本制备程序为低成本快速基因分析集成系统的开发提供了重要环节。此外,同一个过滤器可以大量重复使用,检测到的分析之间交叉污染风险极低,且处理能力不会降低,从而进一步降低了每个分析样本的成本。