Aptamer selection by high-throughput sequencing and informatic analysis

Aptamer selection by high-throughput sequencing and informatic analysis
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DOI:
10.2144/000113786
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发表时间:
2011-12-01
期刊:
影响因子:
2.7
通讯作者:
Scolnick, Jonathan
Scolnick, Jonathan
中科院分区:
工程技术4区
文献类型:
--
作者:
Hoon, Shawn;Zhou, Bin;Scolnick, Jonathan

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传统的适体选择方法需要多轮选择和优化,才能识别与其靶标高亲和力结合的适体。在这里,我们描述了一种只需要一轮正选,然后进行高通量 DNA 测序和信息分析即可选择高亲和力适体的测定方法。该测定方法灵活,需要的人工时间较少,并且可以由适体生物学专业知识最少的实验室使用,以快速选择针对感兴趣靶点的高亲和力适体。该测定已用于成功鉴定与凝血酶结合且解离常数在纳摩尔范围内的适体。
Traditional methods for selecting aptamers require multiple rounds of selection and optimization in order to identify aptamers that bind with high affinity to their targets. Here we describe an assay that requires only one round of positive selection followed by high-throughput DNA sequencing and informatic analysis in order to select high-affinity aptamers. The assay is flexible, requires less hands on time, and can be used by laboratories with minimal expertise in aptamer biology to quickly select high-affinity aptamers to a target of interest. This assay has been utilized to successfully identify aptamers that bind to thrombin with dissociation constants in the nanomolar range.