Comparison of Whole-Cell SELEX Methods for the Identification of Staphylococcus Aureus-Specific DNA Aptamers
Abstract
:1. Introduction
2. Materials and Methods
2.1. Biological and Chemical Materials
2.2. Bacterial Strains
2.3. Preparation of DNA Library
2.4. PCR Optimization
2.5. SELEX Procedures
2.6. Fluorescence-Activated Cell Sorting (FACS)
2.7. Characterization of Binding Parameters of Aptamers to Bacteria
2.8. Aptamer-Binding Assays and Prediction of Candidate Aptamer Sequences
3. Results and Discussion
3.1. Screening of Aptamer Candidates in the Basic Whole-Cell SELEX
Aptamers | Sequences |
---|---|
A2 | ACGGGCGTGGGAGGCAATGCCTTGCTTGTAGGCTTCCCCTGTGCGCG |
A14 | CACACCGCAGCAGTGGGAACGTTTCAGCCATGCAAGCATCACGCCCGT |
A15 | CACGCGCAAACAGATTAACACTCCGCCTAAGTCTGCCGCACGC |
A20 | GCGTGCAGCGGGGGCTGCGCGGTGGAGTGCTGTGGGCG |
B3 | GCGTGCGGAGCCAGGATGGGAGGTCTGTAGGTCTGCGGGGCGTG |
B6 | GCGTGTCGGTGTCTGCCGGGGGATGTGGAGGCTGGGTGTTGCGCG |
B7 | GCGTGGGCGGGCTACCTGGCTAGTACGCCATGATGCCTGCACGCG |
B15 | CACGCGCAAACAGATTAACACTCCGCCTAAGTCTGCCGCACGC |
3.2. Screening of Aptamer Candidates in the Modified Whole-Cell SELEX
3.3. Dissociation Constants for Aptamers
4. Conclusions/Outlook
Supplementary Files
Supplementary File 1Acknowledgments
Author Contributions
Conflicts of Interest
References
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Moon, J.; Kim, G.; Park, S.B.; Lim, J.; Mo, C. Comparison of Whole-Cell SELEX Methods for the Identification of Staphylococcus Aureus-Specific DNA Aptamers. Sensors 2015, 15, 8884-8897. https://doi.org/10.3390/s150408884
Moon J, Kim G, Park SB, Lim J, Mo C. Comparison of Whole-Cell SELEX Methods for the Identification of Staphylococcus Aureus-Specific DNA Aptamers. Sensors. 2015; 15(4):8884-8897. https://doi.org/10.3390/s150408884
Chicago/Turabian StyleMoon, Jihea, Giyoung Kim, Saet Byeol Park, Jongguk Lim, and Changyeun Mo. 2015. "Comparison of Whole-Cell SELEX Methods for the Identification of Staphylococcus Aureus-Specific DNA Aptamers" Sensors 15, no. 4: 8884-8897. https://doi.org/10.3390/s150408884