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Capillary Electrophoresis-Selex Sele...

Alenezi, Fajer Y.

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Capillary Electrophoresis-Selex Selection of ssDNA Aptamers With Affinity for Nicotine.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Capillary Electrophoresis-Selex Selection of ssDNA Aptamers With Affinity for Nicotine./
作者:	Alenezi, Fajer Y.
出版者:	Ann Arbor : ProQuest Dissertations & Theses, : 2023,
面頁冊數:	65 p.
附註:	Source: Masters Abstracts International, Volume: 85-04.
Contained By:	Masters Abstracts International85-04.
標題:	Chemistry. -
電子資源:	https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=30689354
ISBN:	9798380614016

Capillary Electrophoresis-Selex Selection of ssDNA Aptamers With Affinity for Nicotine.
Alenezi, Fajer Y.

Capillary Electrophoresis-Selex Selection of ssDNA Aptamers With Affinity for Nicotine. - Ann Arbor : ProQuest Dissertations & Theses, 2023 - 65 p.

Source: Masters Abstracts International, Volume: 85-04.

Thesis (M.S.)--San Diego State University, 2023.

This item must not be sold to any third party vendors.

The thirdhand smoke (THS) is the contaminated residue left on surfaces, and in dust, persisting in the environment long after smoking has ceased. It has been known to carry almost the same dangers as secondhand smoke and it is more likely to become more toxic when reacting with air pollutants resulting in the formation of tobacco-specific nitrosamines. Although the current detection methods have been successfully able to detect the presence of THS residues, the process is very sensitive, but costly, and inaccessible to average individuals. For this reason, there is a need for rapid, simple testing for THS residue, particularly for rental housing, where children may be unknowingly exposed to these toxins. In recent years, aptamer selection technology has shown a tremendous progress in detecting molecules including toxins, antibiotics, viruses, and proteins. Here, a ssDNA aptamer screening method was proposed to detect nicotine, the most abundant molecule in THS. The identification of a viable ssDNA aptamer was performed with capillary electrophoresis- systematic evolution of ligands by exponential enrichment (CE-SELEX). Prior to selecting an aptamer for the nicotine, the effect of separation buffer, and pressure on EOF were tested to ensure the reproducibility of the ssDNA library collection. The incubation of nicotine with ssDNA library has resulted in the formation of complex peaks. These peaks were collected and amplified by PCR, in the presence of additional magnesium ions to cause some random mutations, and screened again by CE until a sufficiently sensitive and selective aptamer is found. A limited pool of aptamers was obtained under optimized incubation condition of 1:1 molar ratio of nicotine to ssDNA library incubated in 50 mM sodium borate buffer, pH 8.72, at -4 ℃ for 5 days. This was done in four rounds of CE-SELEX with a K_D value of 0.5 μM. By sequencing the fourth aptamer pool through next-generation sequencing (NGS) and identifying the sequences with high selectivity and specificity for nicotine, we can develop a sensitive, robust, and a simple device to detect the presence of THS contaminants.

ISBN: 9798380614016Subjects--Topical Terms:

516420
Chemistry.
Subjects--Index Terms:

Thirdhand smoke

Capillary Electrophoresis-Selex Selection of ssDNA Aptamers With Affinity for Nicotine.
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520 $a The thirdhand smoke (THS) is the contaminated residue left on surfaces, and in dust, persisting in the environment long after smoking has ceased. It has been known to carry almost the same dangers as secondhand smoke and it is more likely to become more toxic when reacting with air pollutants resulting in the formation of tobacco-specific nitrosamines. Although the current detection methods have been successfully able to detect the presence of THS residues, the process is very sensitive, but costly, and inaccessible to average individuals. For this reason, there is a need for rapid, simple testing for THS residue, particularly for rental housing, where children may be unknowingly exposed to these toxins. In recent years, aptamer selection technology has shown a tremendous progress in detecting molecules including toxins, antibiotics, viruses, and proteins. Here, a ssDNA aptamer screening method was proposed to detect nicotine, the most abundant molecule in THS. The identification of a viable ssDNA aptamer was performed with capillary electrophoresis- systematic evolution of ligands by exponential enrichment (CE-SELEX). Prior to selecting an aptamer for the nicotine, the effect of separation buffer, and pressure on EOF were tested to ensure the reproducibility of the ssDNA library collection. The incubation of nicotine with ssDNA library has resulted in the formation of complex peaks. These peaks were collected and amplified by PCR, in the presence of additional magnesium ions to cause some random mutations, and screened again by CE until a sufficiently sensitive and selective aptamer is found. A limited pool of aptamers was obtained under optimized incubation condition of 1:1 molar ratio of nicotine to ssDNA library incubated in 50 mM sodium borate buffer, pH 8.72, at -4 ℃ for 5 days. This was done in four rounds of CE-SELEX with a K_D value of 0.5 μM. By sequencing the fourth aptamer pool through next-generation sequencing (NGS) and identifying the sequences with high selectivity and specificity for nicotine, we can develop a sensitive, robust, and a simple device to detect the presence of THS contaminants.
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