Genome Sequencing Platform, Broad Institute of MIT and Harvard, 320 Charles St, Cambridge, MA 02141, USA.
Genome Biol. 2010;11(2):R15. doi: 10.1186/gb-2010-11-2-r15. Epub 2010 Feb 5.
We present an automated, high throughput library construction process for 454 technology. Sample handling errors and cross-contamination are minimized via end-to-end barcoding of plasticware, along with molecular DNA barcoding of constructs. Automation-friendly magnetic bead-based size selection and cleanup steps have been devised, eliminating major bottlenecks and significant sources of error. Using this methodology, one technician can create 96 sequence-ready 454 libraries in 2 days, a dramatic improvement over the standard method.
我们提出了一种自动化、高通量的 454 技术文库构建方法。通过对塑料制品进行端到端的条码标记以及对构建体进行分子 DNA 条码标记，最大限度地减少了样品处理错误和交叉污染。设计了自动化友好型基于磁珠的大小选择和清洗步骤，消除了主要的瓶颈和重大误差源。使用这种方法，一名技术人员可以在 2 天内创建 96 个序列准备好的 454 文库，这比标准方法有了显著的改进。
