Magnetic-silica nanoshell for extraction of fungal genomic DNA from Rhizopus oryzae

被引:9
作者
Alghuthaymi, Mousa [1 ]
机构
[1] Shaqra Univ, Sci & Humanities Coll, Dept Biol, Alquwayiyah, Saudi Arabia
关键词
Rhizopus; DNA isolation; PCR enhancement; magnetic nanoparticles; NANOPARTICLES; PURIFICATION; BINDING; PCR; AGGREGATION; SEPARATION;
D O I
10.33263/BRIAC102.972976
中图分类号
O69 [应用化学];
学科分类号
081704 ;
摘要
The main objective of the present article describes an established genomic DNA purification technique improved in vitro by using magnetic nanoparticles as a stable phase adsorbent. Stober technique was used to synthesis Fe3O4@SiO3 nanoshell, and described using transmission electron microscopy (TEM) and dynamic light scattering. The quality and quantity of purified DNA were verified by using agarose gel electrophoresis and some PCR strategies such as universal-primed UP-PCR, RAPD patterns. Up to 50 mu g of DNA was obtained from a slight quantity of one hundred mg of fungal mycelium. Fungal DNA produced with the aid of this technique was applied as a fungal DNA template for PCR method to amplify UP-PCR, RAPD provided replicable profiles. DNA purified from fungal mats was 100 % pure enough to generally be used at high concentrations for PCR technique. The end result confirmed that Fe3O4@SiO3 nanoparticles of similar to 140 nm diameter with concentration of 0.4 mg/ml produced premium quality and amount of isolated DNA produced from fungal mats. The obtained DNA was free from any polluting proteins, polysaccharides and coloured dyes. Magnetic nanoparticles (MNP)-mediated genomic DNA extraction is pretty simple (loose from filtration and centrifugation), speedy (30 min), and ecofriendly (not contain any toxic chemical compounds).
引用
收藏
页码:4972 / 4976
页数:5
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