Banér Johan, Isaksson Anders, Waldenström Erik, Jarvius Jonas, Landegren Ulf, Nilsson Mats
The Beijer Laboratory, Department of Genetics and Pathology, Rudbeck Laboratory, SE-751 85 Uppsala, Sweden.
Nucleic Acids Res. 2003 Sep 1;31(17):e103. doi: 10.1093/nar/gng104.
Parallel, highly specific analysis methods are required to take advantage of the extensive information about DNA sequence variation and of expressed sequences. We present a scalable laboratory technique suitable to analyze numerous target sequences in multiplexed assays. Sets of padlock probes were applied to analyze single nucleotide variation directly in total genomic DNA or cDNA for parallel genotyping or gene expression analysis. All reacted probes were then co-amplified and identified by hybridization to a standard tag oligonucleotide array. The technique was illustrated by analyzing normal and pathogenic variation within the Wilson disease-related ATP7B gene, both at the level of DNA and RNA, using allele-specific padlock probes.
需要采用平行、高度特异的分析方法来利用有关DNA序列变异和表达序列的大量信息。我们提出了一种可扩展的实验室技术，适用于在多重分析中分析众多目标序列。使用锁式探针组直接在总基因组DNA或cDNA中分析单核苷酸变异，以进行平行基因分型或基因表达分析。然后将所有反应的探针进行共扩增，并通过与标准标签寡核苷酸阵列杂交来鉴定。通过使用等位基因特异性锁式探针，在DNA和RNA水平分析威尔逊病相关ATP7B基因内的正常和致病变异，对该技术进行了说明。
