基于微珠的寡核苷酸连接单核苷酸多态性基因分型检测方法的改进。
Improvements to bead-based oligonucleotide ligation SNP genotyping assays.
作者信息
Bruse Shannon, Moreau Michael, Azaro Marco, Zimmerman Ray, Brzustowicz Linda
机构信息
Department of Genetics, Rutgers University, Piscataway, NJ 08854, USA.
出版信息
Biotechniques. 2008 Nov;45(5):559-71. doi: 10.2144/000112960.
Abstract
We describe a bead-based, multiplexed, oligonucleotide ligation assay (OLA) performed on the Luminex flow cytometer. Differences between this method and those previously reported include the use of far fewer beads and the use of a universal oligonucleotide for signal detection. These innovations serve to significantly reduce the cost of the assay, while maintaining robustness and accuracy. Comparisons are made between the Luminex OLA and both pyrosequencing and direct sequencing. Experiments to assess conversion rates, call rates, and concordance across technical replicates are also presented.
摘要
我们描述了一种基于磁珠的多重寡核苷酸连接分析（OLA），该分析在Luminex流式细胞仪上进行。此方法与先前报道的方法之间的差异包括使用的磁珠数量少得多，以及使用通用寡核苷酸进行信号检测。这些创新有助于显著降低分析成本，同时保持稳健性和准确性。对Luminex OLA与焦磷酸测序和直接测序进行了比较。还展示了评估技术重复之间的转化率、检出率和一致性的实验。
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