Aptamer-Based Regionally Protected PCR for Protein Detection

Abstract

BACKGROUND: DNA aptamers are single-stranded nucleotide sequences that bind specifically to target molecules. By combining the advantages of PCR for amplifying specific DNA sequences and aptamer technology, we have developed a new strategy to detect target molecules such as proteins.

SELEX tool: a novel and convenient gel-based diffusion method for monitoring of aptamer-target binding

“…A variety of methods for monitoring of SELEX process have been reported. We have evaluated a range of approaches including the methods of EMSA [34], dot blotting, Eastern blotting and target-capture assay [51], quartz crystal microbalance (QCM) analysis [52], qPCR (data not shown), HTS technology (data not shown), and GBDM in our laboratory. We rank the methods.…”

Fluorescent aptamers for detection and treatment of pathogenic bacteria and cancer

“…Combining the attributes of high specificity binding of DNA aptamers to several kinds of target molecules with PCR, low concentration analytes can be readily calculated. This detection assay termed as aptamer-based regionally protected PCR (ARP-PCR) features the complex formation of the DNA aptamer and its target with subsequent DNAse I treatment before amplification such that the unbound aptamer segments are cleaved (Lin & McNatty, 2009). As a result, only the target-bound portion of the DNA aptamer is amplified and sequenced.…”

An aptamer-based sensing platform for luteinising hormone pulsatility measurement

“…The reason I choose nitrocellulose membrane selection is I don't have any protein tag in our recombinant human LH protein for the selection. There is currently no LH aptamer exist in the literature, only hCG peptide aptamer [144] and Ovine FSH α subunit DNA aptamer [145] have been reported. As the fact that our LH DNA aptamer is specific to LH but not FSH so I assumed the binding side of the aptamer might be in the β subunit which gives us more novelty of the project.…”