Wang Jiuhai, Ali Zeeshan, Si Jin, Wang Nianyue, He Nongyue, Li Zhiyang
J Nanosci Nanotechnol. 2017 Jan;17(1):802-06. doi: 10.1166/jnn.2017.12442.
Nucleic acid (NA) extraction from cancer cells is an essential step in molecular oncologic testing. The conventional NA extraction protocols, based on several ultracentrifugation steps, suffer from time-consuming and complex manipulation. Here, a magnetic nanoparticle (MNP) based method for simultaneous extraction of DNA and RNA from cancer cells is described. This MNP based technique has received great attention and significant interest due to its convenient manipulation, low cost and ease for automation. Different factors including lysis buffer, ethanol, MNPs and washing buffers which may affect the yield of nucleic acid were optimized. The average yield of DNA and RNA obtained from 1 mL Hep G2 (˜106 cells) ranged from 9.7 to 14.7 μg with A260/A280 values between 1.68 and 2.01. The isolated DNA and RNA, using this method, were suitable for downstream activities such as PCR and RT-PCR.
从癌细胞中提取核酸是分子肿瘤学检测的关键步骤。基于多次超速离心步骤的传统核酸提取方案存在操作耗时且复杂的问题。在此,描述了一种基于磁性纳米颗粒(MNP)从癌细胞中同时提取DNA和RNA的方法。这种基于MNP的技术因其操作简便、成本低且易于自动化而备受关注。对包括裂解缓冲液、乙醇、MNP和洗涤缓冲液等可能影响核酸产量的不同因素进行了优化。从1 mL Hep G2(约106个细胞)中获得的DNA和RNA的平均产量在9.7至14.7 μg之间,A260/A280值在1.68至2.01之间。使用该方法分离的DNA和RNA适用于下游活动,如PCR和RT-PCR。