Khan Imran, Kaushik Gaurav, Verma Chaitenya, Vashishtha Richa, Kumar Vinay
School of Allied Health Sciences, Sharda University, Greater Noida, Uttar Pradesh, 201310, India.
Department of Biotechnology, Sharda University, Greater Noida, Uttar Pradesh, 201310, India.
Nanotechnol Sci Appl. 2024 Dec 14;17:247-258. doi: 10.2147/NSA.S494613. eCollection 2024.
Efficient extraction of DNA from biological fluids is crucial for applications in molecular biology, forensic science, and clinical diagnostics. However, traditional DNA extraction methods often require costly reagents and lengthy procedures. This study aims to optimize the binding buffer composition for DNA extraction using polyethyleneimine-coated iron oxide nanoparticles (PEI-IONPs), which offer the dual benefits of magnetic separation and high DNA-binding efficiency.
The effects of three key binding buffer components-polyethylene glycol (PEG-6000), sodium chloride (NaCl), and pH-on DNA adsorption efficiency were systematically evaluated. Blood samples were treated with PEI-IONPs under various conditions, and DNA concentration, yield, and purity were quantified. Nanoparticle functionalization was confirmed through characterization, and DNA quality was validated via agarose gel electrophoresis.
The optimized binding buffer composition consisted of a PEG-6000 concentration of 30%, NaCl concentration of 0M, and pH of 4, which yielded the highest DNA concentration (34 ± 1.2 ng/μL), yield (6.8 ± 0.2 μg), and purity (A260/A280 ratio of 1.81). These conditions significantly improved DNA recovery compared to suboptimal buffer compositions.
The findings highlighted the critical role of binding buffer composition in maximizing DNA recovery. The use of optimized PEI-IONPs provided a rapid and efficient method for DNA extraction, supporting its potential for applications in scientific and clinical research. Future studies should explore the robustness of these optimized conditions across diverse biological fluids and extraction settings.
从生物体液中高效提取DNA对于分子生物学、法医学和临床诊断应用至关重要。然而，传统的DNA提取方法通常需要昂贵的试剂和冗长的程序。本研究旨在优化使用聚乙烯亚胺包覆的氧化铁纳米颗粒（PEI-IONPs）进行DNA提取的结合缓冲液组成，该纳米颗粒具有磁分离和高DNA结合效率的双重优势。
系统评估了三种关键结合缓冲液成分——聚乙二醇（PEG-6000）、氯化钠（NaCl）和pH值——对DNA吸附效率的影响。在各种条件下用PEI-IONPs处理血样，并对DNA浓度、产量和纯度进行定量。通过表征确认纳米颗粒功能化，并通过琼脂糖凝胶电泳验证DNA质量。
优化后的结合缓冲液组成包括30%的PEG-6000浓度、0M的NaCl浓度和pH值4，其产生了最高的DNA浓度（34±1.2 ng/μL）、产量（6.8±0.2μg）和纯度（A260/A280比值为1.81）。与次优缓冲液组成相比，这些条件显著提高了DNA回收率。
研究结果突出了结合缓冲液组成在最大化DNA回收率方面的关键作用。使用优化的PEI-IONPs提供了一种快速高效的DNA提取方法，支持其在科学和临床研究中的应用潜力。未来的研究应探索这些优化条件在不同生物体液和提取环境中的稳健性。
