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oai:www.ideals.illinois.edu:2142/42165 2023-07-11 col_2142_5131 col_2142_14770 com_2142_5130 com_2142_3518 com_2142_234

Werth, Charles J. Akki, Spurti Umesh 2013-02-03T19:18:02Z 2013-02-03T19:18:02Z 2015-02-03T11:00:44Z 2012-12 2013-02-03T19:18:02Z 2012-12 17β-estradiol (E2) is a potent estrogen that has been widely documented in water resources. It falls under the category of endocrine disrupting compounds (EDCs), and hence has been included in the EPA’s Unregulated Contaminant Monitoring Rule (UCM3) that will require the monitoring of this compound. Thus, there is an increasing demand to sensitively and selectively detect E2. Conventional detection methods are time consuming and/or expensive. We explore a promising approach which employs the use of DNA aptamers for the detection of E2. DNA aptamers are single stranded DNA molecules which are capable of binding target molecules with high affinity and selectivity. The aim of this project is to develop DNA aptamers that would function as sensors for the detection of this endocrine disrupting steroid hormone. The first step performed was iterative in vitro selections to identify aptamers from a pool of ~ 1014 DNA molecules with the application of appropriate selection pressures. This was achieved by passing the pool over a selection column containing the target immobilized on sepharose beads, followed by eluting the sequences bound to the column with the free target. The resulting pool with the desired sequences was enriched by carrying out polymerase chain reaction (PCR). The potential aptamers obtained were cloned and sequenced, following which they were screened for binding activity using a bead binding assay. The dissociation constant for each aptamer was determined by a DMS chemical probing assay. These aptamers can be incorporated into various real-time in-line sensing platforms to generally and selectively monitor the presence of E2 in environmental samples. They represent an improvement over the one existing aptamer for E2, because some are more selective and marginally more sensitive. Item withdrawn by Mark Zulauf (zulauf@illinois.edu) on 2012-12-11T18:27:13Z Item was in collections: University of Illinois Theses & Dissertations (ID: 1) No. of bitstreams: 2 Akki_Spurti_MS thesis_final.docx: 943282 bytes, checksum: 74299280c10d2ce853e5a17f480e281c (MD5) Akki_Spurti Umesh.pdf: 1215403 bytes, checksum: cbe92d8cf6dfec3e7d20adaf95c3dd76 (MD5) Made available in DSpace on 2013-02-03T19:18:02Z (GMT). No. of bitstreams: 3 Spurti Umesh_Akki.pdf: 1215403 bytes, checksum: cbe92d8cf6dfec3e7d20adaf95c3dd76 (MD5) Akki_Spurti_MS thesis_final.docx: 943282 bytes, checksum: 74299280c10d2ce853e5a17f480e281c (MD5) license.txt: 4064 bytes, checksum: 22a8b309f8e331d08f6a5d4b3c2ce2a3 (MD5) Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Seth Robbins (srobbins@illinois.edu) on 2013-02-03T19:19:05Z Item is restricted until 2015-02-03T19:18:53Z Restriction data tranferred 2014-07-01T11:11:58-05:00 Original Data Group with Access UIUC Users [automated] Release Date: 2015-02-03 13:18:53 UTC Reason: Author requested U of Illinois access only (OA after 2yrs) in Vireo ETD system U of I Only Restriction Lifted for Item 42112 on 2015-02-03T11:00:44Z. http://hdl.handle.net/2142/42165 en Copyright 2012 Spurti Umesh Akki DNA aptamer estradiol Selecting DNA aptamers for 17β-estradiol text Civil and Environmental Engineering 1251 Environmental Engineering in Civil Engineering 0231 University of Illinois at Urbana-Champaign Thesis M.S. MS:Env Engr Civil Engr -UIUC 10KS0231MS