A scalable, fully automated process for construction of sequence-ready barcoded libraries for 454.

A scalable, fully automated process for construction of sequence-ready barcoded libraries for 454.
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DOI:
10.1186/gb-2010-11-2-r15
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发表时间:
2010
期刊:
影响因子:
12.3
通讯作者:
Nicol R
Nicol R
中科院分区:
生物学1区
文献类型:
--
作者:
Lennon NJ;Lintner RE;Anderson S;Alvarez P;Barry A;Brockman W;Daza R;Erlich RL;Giannoukos G;Green L;Hollinger A;Hoover CA;Jaffe DB;Juhn F;McCarthy D;Perrin D;Ponchner K;Powers TL;Rizzolo K;Robbins D;Ryan E;Russ C;Sparrow T;Stalker J;Steelman S;Weiand M;Zimmer A;Henn MR;Nusbaum C;Nicol R

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构建454测序库的自动化方法显着降低了所需的成本和时间。我们提出了一个自动化的,高通量的454技术库建设过程。通过塑料器皿的端到端条形码沿着构建体的分子DNA条形码,最大限度地减少了样品处理错误和交叉污染。已经设计出了自动化友好的基于磁珠的尺寸选择和清理步骤,消除了主要瓶颈和重大错误来源。使用这种方法,一名技术人员可以在2天内创建96个测序就绪的454文库,这比标准方法有了显着的改进。
An automated method for constructing libraries for 454 sequencing significantly reduces the cost and time required. 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.
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