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Please use this identifier to cite or link to this item: http://repository.iitr.ac.in/handle/123456789/20163
Title: A reliable, quick and universally applicable method for monitoring aptamer SELEX progress
Authors: Baba S.A.
Jain S.
Navani, Naveen Kumar
Published in: Gene
Abstract: Nucleic acid aptamers for biosensing are developed from a complex ssDNA library through Systematic Evolution of Ligands by Exponential Enrichment (SELEX) process. Monitoring of SELEX process is crucial for generating high-affinity aptamers. Extant methods for monitoring aptamer selection are either arduous or give false-positive signals, which adversely impact the outcome of selection. We describe a colorimetric, simple and cost-effective, novel method to monitor the progress of in vitro selections. The power of rolling circle amplification (RCA) and inherent Horse Radish Peroxidase (HRP)-mimicking activity of G-quadruplex/hemin DNAzyme were employed to produce a colorimetric signal. A unique extension of DNA population at 3ʹ-OH end by PCR generated concatenated repeats by rolling circle amplification (RCA) reaction. Oxidation of substrate ABTS (2, 2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) in presence of H2O2 and hemin cofactor produced colorimetric signal. Analysis of the signal generated by the DNA pool bound to their target provided a quantitative measurement of SELEX. We demonstrate the reproducibility and accuracy of the method by evaluating the progress of two discrete selections. © 2021 Elsevier B.V.
Citation: Gene, 774
URI: https://doi.org/10.1016/j.gene.2021.145416
http://repository.iitr.ac.in/handle/123456789/20163
Issue Date: 2021
Publisher: Elsevier B.V.
Keywords: ABTS
Aptamer
Colorimetric detection
DNAzyme G-quadruplexes
Rolling Circle Amplification reaction
SELEX
ISSN: 3781119
Author Scopus IDs: 57210897210
57213028727
6507073924
Author Affiliations: Baba, S.A., Chemical Biology Lab, Department of Biotechnology, Indian Institute of Technology, Roorkee, 247667, India
Jain, S., Chemical Biology Lab, Department of Biotechnology, Indian Institute of Technology, Roorkee, 247667, India
Navani, N.K., Chemical Biology Lab, Department of Biotechnology, Indian Institute of Technology, Roorkee, 247667, India
Funding Details: This work was supported by funding from Department of Biotechnology, Government of India via Grant No.BT/P21783/NNT/28/1219/2017. The sequences used in the study have been deposited in Indian Patent Office as part of the Indian Patent titled “Single-stranded nucleotide sequences specific for binding to Acinetobacter baumannii and uses thereof” with application No. 201911016947 Dated: 29/04/2019. /P21783/NNT/28/1219/2017
Corresponding Author: Navani, N.K.; Chemical Biology Lab, India; email: naveen.navani@bt.iitr.ac.in
Appears in Collections:Journal Publications [BT]

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