Bau Stephan, Schracke Nadine, Kränzle Marcel, Wu Haiguo, Stähler Peer F, Hoheisel Jörg D, Beier Markus, Summerer Daniel
febit biomed gmbh, Im Neuenheimer Feld 519, 69120, Heidelberg, Germany.
Anal Bioanal Chem. 2009 Jan;393(1):171-5. doi: 10.1007/s00216-008-2460-7. Epub 2008 Oct 29.
We report a flexible method for selective capture of sequence fragments from complex, eukaryotic genome libraries for next-generation sequencing based on hybridization to DNA microarrays. Using microfluidic array architecture and integrated hardware, the process is amenable to complete automation and does not introduce amplification steps into the standard library preparation workflow, thereby avoiding bias of sequence distribution and fragment lengths. We captured a discontiguous human genomic target region of 185 kb using a tiling design with 50mer probes. Analysis by high-throughput sequencing using an Illumina/Solexa 1G Genome Analyzer revealed 2150-fold enrichment with mean per base coverage between 4.6 and 107.5-fold for the individual target regions. This method represents a flexible and cost-effective approach for large-scale resequencing of complex genomes.
我们报告了一种灵活的方法，可基于与DNA微阵列杂交，从复杂的真核基因组文库中选择性捕获序列片段，用于下一代测序。利用微流控阵列架构和集成硬件，该过程适合完全自动化，且不会在标准文库制备工作流程中引入扩增步骤，从而避免了序列分布和片段长度的偏差。我们使用50聚体探针的平铺设计捕获了一个185 kb的不连续人类基因组目标区域。使用Illumina/Solexa 1G基因组分析仪进行的高通量测序分析显示，单个目标区域的富集倍数为2150倍，平均每碱基覆盖度在4.6至107.5倍之间。该方法代表了一种用于复杂基因组大规模重测序的灵活且经济高效的方法。
