Simultaneous extraction of DNA and RNA from Escherichia coli BL 21 based on silica-coated magnetic nanoparticles  

作　　者：Jiuhai Wang Zeeshan Ali Nianyue Wang Wenbiao Liang Hongna Liu Fu Li Haowen Yang Lei He Libo Nie Nongyue He Zhiyang Li 

机构地区：[1]State Key Laboratory of Bioelectronics [2]School of Biological Science and Medical Engineering, Southeast University [3]Clinical Laboratory, the Second Hospital of Nanjing [4]Jiangsu Province Blood Center [5]Economical Forest Cultivation and Utilization of 2011 Collaborative Innovation Center in Hunan Province [6]Hunan Key Laboratory of Green Packaging and Application of Biological Nanotechnology [7]Hunan University of Technology [8]Department of Laboratory Medicine, the Second Affiliated Hospital of Nanjing Medical University

出　　处：《Science China Chemistry》2015年第11期1774-1778,共5页中国科学（化学英文版）

基　　金：supported by the National Basic Research Program of China(2014CB744501);the National High Technology Research and Development Program of China(2012AA022703);the National Key Special Science Program(2013ZX10004103-002);the National Natural Science Foundation of China(61201033,21205013,61271056,61527806);Projects of Development of Science and Medical Technology(201208038);Projects of Health Ministry of Nanjing(ZKX12038);the Clinical Science and Technology Special Projects in Jiangsu Province(BL2012067,BL2014094);the Talents Planning of Six Summit Fields of Jiangsu Province(2013-WSN-056);China Postdoctoral Science Foundation Funded Project(2014M551491,2015T80487);Jiangsu Planned Projects for Postdoctoral Research Funds(1302007A);the Economical Forest Cultivation and Utilization of 2011 Collaborative Innovation Center in Hunan Province

摘　　要：The extraction of nucleic acid is recognized as one of the most essential steps in molecular biology for initiating other downstream applications such as sequencing, amplification, hybridization, and cloning. Many commercial kits and methods are currently available that allow the extraction of only one type of nucleic acids-DNA or RNA. However, in parallel clinical detection of several diseases, a method for simultaneous extraction of both DNA and RNA from the same source is needed in such cases. In this study, a method for simultaneous extraction of DNA and RNA from bacteria based on magnetic nanoparticles(MNPs) was described. Lysis buffers were prepared to help the nucleic acid released and adsorbed to MNPs. Then, two washing buffers were used to remove the contamination of proteins and carbohydrates. The nucleic acids were finally eluted by Deoxyribonuclease(DNase) and Ribonucleases(RNase) free water. Different factors which might affect the purification of the nucleic acid were investigated, and the quantity and quality parameters of the nucleic acid were also recorded. The DNA and RNA extracted from bacteria were then respectively subjected to polymerase chain reaction(PCR) and reverse transcription PCR(RT-PCR) to further confirm its quality. The results indicated that our method can be successfully used to simultaneously extract DNA and RNA from bacteria.The extraction of nucleic acid is recognized as one of the most essential steps in molecular biology for initiating other downstream applications such as sequencing, amplification, hybridization, and cloning. Many commercial kits and methods are currently available that allow the extraction of only one type of nucleic acids-DNA or RNA. However, in parallel clinical detection of several diseases, a method for simultaneous extraction of both DNA and RNA from the same source is needed in such cases. In this study, a method for simultaneous extraction of DNA and RNA from bacteria based on magnetic nanoparticles （MNPs） was described. Lysis buffers were prepared to help the nucleic acid released and adsorbed to MNPs. Then, two washing buffers were used to remove the contamination of proteins and carbohydrates. The nucleic acids were finally eluted by Deoxyribonuclease （DNase） and Ribonucleases （RNase） free water. Different factors which might affect the purification of the nucleic acid were investigated, and the quantity and quality parameters of the nucleic acid were also recorded. The DNA and RNA extracted from bacteria were then respectively subjected to polymerase chain reaction （PCR） and reverse transcription PCR （RT-PCR） to further confirm its quality. The results indicated that our method can be successfully used to simultaneously extract DNA and RNA from bacteria.

关 键 词：nucleic acids simultaneous extraction magnetic nanoparticles （MNPs） BACTERIA 

分 类 号：O629.74[理学—有机化学] TB383.1[理学—化学]