Aptamer selection by high-throughput sequencing and informatic analysis

被引:70
|
作者
Hoon, Shawn [2 ]
Zhou, Bin
Janda, Kim D.
Brenner, Sydney [1 ,2 ]
Scolnick, Jonathan [1 ,2 ]
机构
[1] Scripps Res Inst, Dept Physiol Chem, La Jolla, CA 92037 USA
[2] Agcy Sci Technol & Res, Mol Engn Lab, Singapore, Singapore
关键词
HUMAN THROMBIN; RNA MOLECULES; BIND; LIGANDS;
D O I
10.2144/000113786
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:413 / 416
页数:4
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