Magnetic nano fluids for isolation of genomic DNA and total RNA from various prokaryote and eukaryote cells
Magnetic nano fluids for isolation of genomic DNA and total RNA from various prokaryote and eukaryote cells
复制标题
DOI：
10.1016/j.jchromb.2018.10.006
复制
发表时间：
2018-12-01
影响因子：
3
通讯作者：
Nematzadeh, Ghorban Ali
中科院分区：
文献类型：
--
作者：
Ghahari, Sajjad;Ghahari, Somayeh;Nematzadeh, Ghorban Ali
从各种细胞中正确分离核酸是许多生物化学和诊断过程如克隆、测序、复制、杂交和互补DNA（cDNA）合成之前的重要预备步骤。本研究利用Tween 20和油酸对磁性纳米粒子（MNF）的顺磁性和生物相容性，将其用于原核和真核细胞基因组DNA（gDNA）和总RNA的提取。通过琼脂糖凝胶电泳、酶切和聚合酶链反应（PCR）技术证明提取的gDNA和总RNA的量和准确性。根据紫外可见分光光度数据和琼脂糖凝胶上观察到的gDNA和核糖体RNA（rRNA）条带，结果表明该纳米试剂盒的提取可以与现有的用于纯化核酸的方法（例如基于使用十六烷基三甲基溴化铵（CTAB）和苯酚-氯仿法的纯化）相媲美。颗粒的表征将它们定义为直径类似于34.85 nm，并表现出高饱和磁化强度（28 emu/g）。该方法的优点是可以消除萃取液中的苯酚和氯仿等有害试剂，用有机油代替二氧化硅等无机涂层，缩短反应时间。因此，根据核酸纯化途径中的挑战，这些试剂盒的使用可以是显著的。
The correct isolation of nucleic acid from various cells is an important preliminary step before many biochemical and diagnostic processes such as cloning, sequencing, replication, hybridization, and complementary DNA (cDNA) synthesis. In this study, the coated magnetic nanoparticles (MNFs) with Tween 20 and oleic acid because of paramagnetic and bio-compatibility properties used in the extractions of genomic DNA (gDNA) and total RNA from prokaryote and eukaryote cells. The amount and accuracy of gDNA and total RNA extracted were proved via agarose gel electrophoresis, digestion and polymerase chain reaction (PCR) techniques. According to UV-Vis spectrophotometry data and gDNA and ribosomal RNA (rRNA) bands observed on the agarose gel, the results showed that extraction of this nano-kit can be comparable with the existing methods used to purifying nucleic acids such as purification based on the use of Cetyltrimethylammonium bromide (CTAB) and phenol-chloroform methods. Characterization of the particles defines them to be similar to 34.85 nm in diameter and exhibiting high saturation magnetization (28 emu/g). Elimination of hazardous reagents such as phenol and chloroform from extraction solutions, the replacement for inorganic coating such as silica with organic oil, and reduction of reaction time are some advantages of this method. Therefore, according to the challenges in the nucleic acid purification pathway, the use of these kits can be remarkable.