Real-Time PCR-Coupled CE-SELEX for DNA Aptamer Selection
Joint Authors
Pai, Rekha B.
Ruff, Patrick
Storici, Francesca
Source
Issue
Vol. 2012, Issue 2012 (31 Dec. 2012), pp.1-9, 9 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2012-08-08
Country of Publication
Egypt
No. of Pages
9
Main Subjects
Natural & Life Sciences (Multidisciplinary)
Abstract EN
Aptamers are short nucleic acid or peptide sequences capable of binding to a target molecule with high specificity and affinity.
Also known as “artificial antibodies,” aptamers provide many advantages over antibodies.
One of the major hurdles to aptamer isolation is the initial time and effort needed for selection.
The systematic evolution of ligands by exponential enrichment (SELEX) is the traditional procedure for generating aptamers, but this process is lengthy and requires a large quantity of target and starting aptamer library.
A relatively new procedure for generating aptamers using capillary electrophoresis (CE), known as CE-SELEX, is faster and more efficient than SELEX but requires laser-induced fluorescence (LIF) to detect the aptamer-target complexes.
Here, we implemented an alternative system without LIF using real-time- (RT-) PCR to indirectly measure aptamer-target complexes.
In three rounds of selection, as opposed to ten or more rounds common in SELEX protocols, a specific aptamer for bovine serum albumin (BSA) was obtained.
The specificity of the aptamer to BSA was confirmed by electrophoretic mobility shift assays (EMSAs), an unlabeled competitor assay, and by a supershift assay.
The system used here provides a cost effective and a highly efficient means of generating aptamers.
American Psychological Association (APA)
Ruff, Patrick& Pai, Rekha B.& Storici, Francesca. 2012. Real-Time PCR-Coupled CE-SELEX for DNA Aptamer Selection. ISRN Molecular Biology،Vol. 2012, no. 2012, pp.1-9.
https://search.emarefa.net/detail/BIM-509808
Modern Language Association (MLA)
Ruff, Patrick…[et al.]. Real-Time PCR-Coupled CE-SELEX for DNA Aptamer Selection. ISRN Molecular Biology No. 2012 (2012), pp.1-9.
https://search.emarefa.net/detail/BIM-509808
American Medical Association (AMA)
Ruff, Patrick& Pai, Rekha B.& Storici, Francesca. Real-Time PCR-Coupled CE-SELEX for DNA Aptamer Selection. ISRN Molecular Biology. 2012. Vol. 2012, no. 2012, pp.1-9.
https://search.emarefa.net/detail/BIM-509808
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-509808