Gunderson Kevin L, Kruglyak Semyon, Graige Michael S, Garcia Francisco, Kermani Bahram G, Zhao Chanfeng, Che Diping, Dickinson Todd, Wickham Eliza, Bierle Jim, Doucet Dennis, Milewski Monika, Yang Robert, Siegmund Chris, Haas Juergen, Zhou Lixin, Oliphant Arnold, Fan Jian-Bing, Barnard Steven, Chee Mark S
Illumina, Inc., San Diego, California 92121, USA.
Genome Res. 2004 May;14(5):870-7. doi: 10.1101/gr.2255804. Epub 2004 Apr 12.
We have developed a simple and efficient algorithm to identify each member of a large collection of DNA-linked objects through the use of hybridization, and have applied it to the manufacture of randomly assembled arrays of beads in wells. Once the algorithm has been used to determine the identity of each bead, the microarray can be used in a wide variety of applications, including single nucleotide polymorphism genotyping and gene expression profiling. The algorithm requires only a few labels and several sequential hybridizations to identify thousands of different DNA sequences with great accuracy. We have decoded tens of thousands of arrays, each with 1520 sequences represented at approximately 30-fold redundancy by up to approximately 50,000 beads, with a median error rate of <1 x 10(-4) per bead. The approach makes use of error checking codes and provides, for the first time, a direct functional quality control of every element of each array that is manufactured. The algorithm can be applied to any spatially fixed collection of objects or molecules that are associated with specific DNA sequences.
我们开发了一种简单高效的算法,通过杂交来识别大量与DNA相连的物体中的每一个成员,并将其应用于制造孔中随机组装的珠子阵列。一旦该算法用于确定每个珠子的身份,微阵列就可用于多种应用,包括单核苷酸多态性基因分型和基因表达谱分析。该算法仅需少量标签和几次连续杂交,就能以极高的准确性识别数千种不同的DNA序列。我们已经解码了数万个阵列,每个阵列有1520个序列,由多达约50000个珠子以约30倍冗余度表示,每个珠子的中位错误率<1×10⁻⁴。该方法利用了错误校验码,并首次对制造的每个阵列的每个元件提供了直接的功能质量控制。该算法可应用于与特定DNA序列相关的任何空间固定的物体或分子集合。