研究目的
To overcome the limitations of current biophysical methods for high-throughput assessment of secondary structures adopted by DNA oligonucleotides by developing a fluorescence sensor array for rapid screening and classification of DNA sequences based on their secondary structure.
研究成果
The developed fluorescence sensor array, combined with multivariate analysis, provides a novel and efficient method for the high-throughput assessment of DNA secondary structures. It successfully discriminates among different topological classes of G4-DNA, single-stranded, and duplex oligonucleotides, offering a valuable tool for the discovery of particular motifs and the classification of DNA sequences based on their secondary structure.
研究不足
The approach is limited to relatively short synthetic oligonucleotides and may not be directly applicable to longer DNA sequences or complex mixtures without further optimization.
1：Experimental Design and Method Selection:
The study utilized a fluorescence sensor array composed of commercially available dyes for the assessment of DNA secondary structures. Multivariate analysis tools (PCA and LDA) were employed for data analysis.
2：Sample Selection and Data Sources:
A set of 34 DNA samples with well-established conformations was used to represent the five groups of interest: parallel, anti-parallel, and hybrid G4s, single-stranded, and double-stranded structures.
3：List of Experimental Equipment and Materials:
Fluorescence emission was recorded using a microplate reader (BMG FluoStar Omega), and CD spectra were recorded with a Jasco J-1500 spectropolarimeter.
4：Experimental Procedures and Operational Workflow:
The assay was performed in a 96-well plate format, suitable for testing small amounts of DNA. Fluorescence emission data were normalized and subjected to multivariate analysis.
5：Data Analysis Methods:
Principal Component Analysis (PCA) and Linear Discriminant Analysis (LDA) were used to classify DNA samples based on their secondary structure.
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