Laboratory Medicine Center, Lanzhou University Second Hospital, 730030, Lanzhou, China.
Center of Eco-material and Green Chemistry, Chinese Academy of Sciences, Lanzhou, 730000, PR China.
Anal Biochem. 2019 Aug 15;579:9-17. doi: 10.1016/j.ab.2019.05.004. Epub 2019 May 9.
A polydopamine magnetic composite (PDA@FeO) was prepared for the extraction of human genomic DNA and characterized by transmission electron microscopy, X-ray diffraction, FT-IR spectrometer, zeta potential and vibrating sample magnetometry. PDA@FeO based on magnetic solid phase extraction (MSPE) method have highly efficient capture of genomic deoxyribonucleic acid (DNA)and gene fragments ranging from about 100 bp to 200 bp. Compared with commercial beads (Shenggong, China) and spin column nucleic acid extraction kit (Tiangen, China), the PDA coated magnetic nanoparticles display superior genomic DNA extraction capacity (116 mg/g) and yield (90.2%). The isolation protocol used the solutions (composed of PEG and NaCl) with a specific pH for the binding and release of DNA. The procedure can be attributed to the charge switch of amino and hydroxyl groups on surface of the magnetic particle. The extracted DNA with high quality (A260/A280 = 1.82 ± 0.04) can be directly used as template for polymerase chain reaction (PCR) followed by agarose gel electrophoresis. The results showed the new composite to be an ideal adsorbent for separation of DNA which had the advantage of its low cost, high extraction capacity and yield.
一种聚多巴胺磁性复合材料(PDA@FeO)被制备用于提取人类基因组 DNA,并通过透射电子显微镜、X 射线衍射、傅里叶变换红外光谱仪、Zeta 电位和振动样品磁强计进行了表征。基于磁性固相萃取(MSPE)方法的 PDA@FeO 能够高效地捕获基因组脱氧核糖核酸(DNA)和基因片段,大小从约 100bp 到 200bp 不等。与商业珠粒(中国申工)和离心柱核酸提取试剂盒(中国天根)相比,涂覆有聚多巴胺的磁性纳米颗粒显示出优异的基因组 DNA 提取能力(116mg/g)和产率(90.2%)。所使用的分离方案采用了具有特定 pH 的 PEG 和 NaCl 溶液用于 DNA 的结合和释放。该过程可归因于磁性颗粒表面上的氨基和羟基基团的电荷转换。提取的 DNA 具有高质量(A260/A280=1.82±0.04),可直接用作聚合酶链反应(PCR)的模板,随后进行琼脂糖凝胶电泳。结果表明,这种新型复合材料是一种理想的 DNA 分离吸附剂,具有成本低、提取容量和产率高的优点。