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用于从各种原核生物和真核生物细胞中分离基因组 DNA 和总 RNA 的磁性纳米流体。

Magnetic nano fluids for isolation of genomic DNA and total RNA from various prokaryote and eukaryote cells.

机构信息

Genetics and Agricultural Biotechnology Institute of Tabarestan (GABIT), Sari Agricultural Sciences and Natural Resources University, Iran.

Genetics and Agricultural Biotechnology Institute of Tabarestan (GABIT), Sari Agricultural Sciences and Natural Resources University, Iran.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Dec 1;1102-1103:125-134. doi: 10.1016/j.jchromb.2018.10.006. Epub 2018 Oct 8.

Abstract

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 ~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.

摘要

从各种细胞中正确分离核酸是许多生化和诊断过程的重要初步步骤,如克隆、测序、复制、杂交和互补 DNA(cDNA)合成。在这项研究中,由于顺磁性和生物相容性,包覆有吐温 20 和油酸的磁性纳米粒子(MNFs)用于从原核和真核细胞中提取基因组 DNA(gDNA)和总 RNA。通过琼脂糖凝胶电泳、消化和聚合酶链反应(PCR)技术证明了提取的 gDNA 和总 RNA 的量和准确性。根据 UV-Vis 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方法可与现有的核酸纯化方法相媲美,如基于十六烷基三甲基溴化铵(CTAB)和酚氯仿法的纯化。对颗粒的表征表明,它们的直径约为 34.85nm,并表现出高饱和磁化强度(28emu/g)。从提取溶液中去除危险试剂(如苯酚和氯仿)、用有机油代替无机涂层(如二氧化硅)以及减少反应时间是该方法的一些优点。因此,根据核酸纯化途径的挑战,这些试剂盒的使用可能非常显著。

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