Date of Award
Master of Science
Jared L Anderson
The selective extraction and accumulation of specific deoxyribonucleic acid (DNA) fragments is required for targeted nucleic acid analysis in order to minimize interferences from samples with abundant heterogeneous sequences. In addition, it is essential to isolate and preserve nucleic acid from biological sample containing numerous compounds such as endonucleases prior to sample analysis. Very recently, ionic liquid (IL) and magnetic ionic liquid (MIL) based materials have shown significant promise in the area of biological sample preparation and separation. In this thesis, various types of ILs and MILs are exhaustively descripted and utilized for two applications: 1) sequence-specific extraction of DNA and 2) RNA preservation.
Functionalized oligonucleotide probes named as ion-tagged oligonucleotides (ITOs) and disubstituted ion-tagged oligonucleotides (DTOs) that hybridize with complementary DNA targets can be subsequently captured by a hydrophobic MIL support. The ITO- and DTO-MIL system is investigated for sequence-specific extraction of DNA in a relatively low concentration within multiple sample matrices such as blood and plasma. This particle- and aggregation-free extraction method is highly potential and beneficial for analysis based on microfluidic devices.
In addition, RNA samples obtained from yeast cells can be extracted and preserved by several MILs simultaneously. RNA remains the high quantity and structural integrity within MILs at room temperature. Moreover, specific MILs can be used to reduce ribonuclease A (RNase A) degradation of RNA. This RNA preservation method reveals multiple advantages including energy-saving and equipment-independent which makes it applicable to in-field RNA sample preparation.
Zhu, Chenghui, "Magnetic ionic liquids in applications of nucleic acid analysis: Sequence-specific extraction of DNA and preservation solvents of RNA" (2020). Graduate Theses and Dissertations. 18262.