Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire CB10 1SA, UK.
Electrophoresis. 2012 Dec;33(23):3521-8. doi: 10.1002/elps.201200128. Epub 2012 Nov 12.
Size selection can be a critical step in preparation of next-generation sequencing libraries. Traditional methods employing gel electrophoresis lack reproducibility, are labour intensive, do not scale well and employ hazardous interchelating dyes. In a high-throughput setting, solid-phase reversible immobilisation beads are commonly used for size-selection, but result in quite a broad fragment size range. We have evaluated and optimised the use of two semi-automated preparative DNA electrophoresis systems, the Caliper Labchip XT and the Sage Science Pippin Prep, for size selection of Illumina sequencing libraries.
大小选择是制备下一代测序文库的关键步骤。传统的凝胶电泳方法缺乏重现性,劳动强度大,不易扩展,并且使用危险的螯合染料。在高通量环境中,通常使用固相可逆固定化珠进行大小选择,但会导致相当宽的片段大小范围。我们已经评估和优化了两种半自动化制备性 DNA 电泳系统( Caliper Labchip XT 和 Sage Science Pippin Prep )在 Illumina 测序文库大小选择中的应用。