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SSCP screening of individual aptamers
L V Gening1, S A Klincheva, A S Gusev
1Institute of Molecular Genetics, Russian Academy of Sciences, Moscow. geni@img.ras.ru
Biotechniques
|October 30, 2001
Summary
This study introduces a novel method for aptamer selection analysis, adapting the single-strand conformation polymorphism (SSCP) technique. This approach efficiently groups identical aptamer sequences, reducing sequencing needs and enabling rapid identification of specific aptamers.
Area of Science:
- Molecular Biology
- Biotechnology
- Nucleic Acid Chemistry
Background:
- Aptamers are nucleic acid molecules selected for specific binding properties through in vitro methods.
- The Systematic Evolution of Ligands by Exponential Enrichment (SELEX) is a common method for aptamer discovery.
- Analyzing sequences of selected aptamers is crucial but can be labor-intensive.
Purpose of the Study:
- To present a novel method for classifying aptamer sequences obtained from SELEX.
- To reduce the sequencing efforts required in aptamer selection.
- To enable rapid isolation and identification of specific aptamers from complex mixtures.
Main Methods:
- Adaptation of the single-strand conformation polymorphism (SSCP) technique for aptamer sequence analysis.
- Classification of selected aptamer sequences into identity groups.
- Application to human angiotensin I-specific aptamers.
Main Results:
- The SSCP-based method effectively classifies individual aptamer sequences into distinct identity groups.
- This approach significantly minimizes the need for extensive sequencing.
- Rapid isolation and identification of aptamers from a mixture were demonstrated.
Conclusions:
- The adapted SSCP method is an efficient tool for analyzing aptamer pools generated by SELEX.
- This technique streamlines the aptamer selection process by reducing sequencing workload.
- The method facilitates faster identification of high-affinity aptamers.