Smirnov Denis A, Burdick Josh T, Morley Michael, Cheung Vivian G
Department of Genetics, University of Pennsylvania, Philadelphia 19104-4318, USA.
Genes Chromosomes Cancer. 2004 May;40(1):72-7. doi: 10.1002/gcc.20015.
Comparative genomic hybridization (CGH) to metaphase chromosomes is a method for genome-wide detection of chromosomal aberrations in DNA samples. Recent advances in microarray technology have improved CGH by replacing metaphase chromosomes with a collection of mapped genomic clones placed on glass slides. However, it is quite expensive and labor-intensive to prepare DNA from the genomic clones for use in constructing genomic microarrays. Here we used strand-displacement rolling circle amplification (RCA) to manufacture whole-genome microarrays by using a collection of about 4,500 mapped RPCI-11 BAC clones that cover the human genome at approximately a 1-Mb resolution. These genomic microarrays detected all major chromosomal aberrations in cancer cells lines and in cell lines with aneuploidy. In this article, we discuss the advantages of using RCA for the manufacturing of large genomic microarrays.
对中期染色体进行比较基因组杂交（CGH）是一种在DNA样本中进行全基因组染色体畸变检测的方法。微阵列技术的最新进展改进了CGH，用放置在载玻片上的一组已定位基因组克隆取代了中期染色体。然而，从基因组克隆中制备用于构建基因组微阵列的DNA成本很高且劳动强度大。在这里，我们使用链置换滚环扩增（RCA），通过使用一组约4500个已定位的RPCI-11 BAC克隆制造全基因组微阵列，这些克隆以大约1 Mb的分辨率覆盖人类基因组。这些基因组微阵列检测到了癌细胞系和非整倍体细胞系中的所有主要染色体畸变。在本文中，我们讨论了使用RCA制造大型基因组微阵列的优点。
