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Please use this identifier to cite or link to this item: http://repository.iitr.ac.in/handle/123456789/20163
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dc.contributor.authorBaba S.A.-
dc.contributor.authorJain S.-
dc.contributor.authorNavani, Naveen Kumar-
dc.date.accessioned2022-02-17T11:27:56Z-
dc.date.available2022-02-17T11:27:56Z-
dc.date.issued2021-
dc.identifier.citationGene, 774-
dc.identifier.issn3781119-
dc.identifier.other33444681-
dc.identifier.urihttps://doi.org/10.1016/j.gene.2021.145416-
dc.identifier.urihttp://repository.iitr.ac.in/handle/123456789/20163-
dc.description.abstractNucleic 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.-
dc.language.isoen_US-
dc.publisherElsevier B.V.-
dc.relation.ispartofGene-
dc.subjectABTS-
dc.subjectAptamer-
dc.subjectColorimetric detection-
dc.subjectDNAzyme G-quadruplexes-
dc.subjectRolling Circle Amplification reaction-
dc.subjectSELEX-
dc.titleA reliable, quick and universally applicable method for monitoring aptamer SELEX progress-
dc.typeArticle-
dc.scopusid57210897210-
dc.scopusid57213028727-
dc.scopusid6507073924-
dc.affiliationBaba, S.A., Chemical Biology Lab, Department of Biotechnology, Indian Institute of Technology, Roorkee, 247667, India-
dc.affiliationJain, S., Chemical Biology Lab, Department of Biotechnology, Indian Institute of Technology, Roorkee, 247667, India-
dc.affiliationNavani, N.K., Chemical Biology Lab, Department of Biotechnology, Indian Institute of Technology, Roorkee, 247667, India-
dc.description.fundingThis 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-
dc.description.correspondingauthorNavani, N.K.; Chemical Biology Lab, India; email: naveen.navani@bt.iitr.ac.in-
Appears in Collections:Journal Publications [BT]

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