Production and processing of aptamer microarrays
Production and processing of aptamer microarrays
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DOI：
10.1016/j.ymeth.2005.05.009
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发表时间：
2005-09-01
期刊：
影响因子：
4.8
通讯作者：
Ellington, AD
中科院分区：
文献类型：
--
作者：
Collett, JR;Cho, EJ;Ellington, AD
适体是针对其特异性结合特性在体外选择的核酸种类。我们描述了适体微阵列的生产和处理方法，包括高通量的详细程序，5' RNA生物素化适体的酶促合成，并将它们排列到链霉亲和素包被的载玻片上。还提供了用于处理适体微阵列的方法，包括封闭、洗涤、干燥和扫描。显示了荧光标记的靶蛋白的特异性捕获的实例，所述靶蛋白单独在结合缓冲液中或与来自细胞裂解物的标记的细胞内蛋白竞争。考虑到生产多重适体芯片所涉及的挑战，从不同的文献来源的适体，并开发标准化的方法来表征用于生物传感器的捕获试剂的性能。(c)2005年爱思唯尔公司All rights reserved.
Aptamers are nucleic acid species that are selected in vitro for their specific binding properties. We describe methods for the production and processing of aptamer microarrays, including detailed procedures for the high-throughput, enzymatic synthesis of 5' RNA biotinylated aptamers and for arraying them onto streptavidin-coated glass slides. Also presented are methods for processing the aptamer microarrays, including blocking, washing, drying, and scanning. Examples are shown for the specific capture of fluorescently labeled target proteins either alone in binding buffer or in competition with labeled intracellular proteins from cell lysates. Consideration is given to the challenges involved in producing multiplex aptamer chips composed of aptamers taken from disparate literature sources, and to the development of standardized methods for characterizing the performance of capture reagents used in biosensors. (c) 2005 Elsevier Inc. All rights reserved.