4.7 Article

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

Journal

SCIENCE CHINA-CHEMISTRY
Volume 58, Issue 11, Pages 1774-1778

Publisher

SCIENCE PRESS
DOI: 10.1007/s11426-015-5483-x

Keywords

nucleic acids; simultaneous extraction; magnetic nanoparticles (MNPs); bacteria

Funding

  1. National Basic Research Program of China [2014CB744501]
  2. National High Technology Research and Development Program of China [2012AA022703]
  3. National Key Special Science Program [2013ZX10004103-002]
  4. National Natural Science Foundation of China [61201033, 21205013, 61271056, 61527806]
  5. Projects of Development of Science and Medical Technology [201208038]
  6. Health Ministry of Nanjing [ZKX12038]
  7. Clinical Science and Technology Special Projects in Jiangsu Province [BL2014094, BL2012067]
  8. Talents Planning of Six Summit Fields of Jiangsu Province [2013-WSN-056]
  9. China Postdoctoral Science Foundation Funded Project [2014M551491, 2015T80487]
  10. Jiangsu Planned Projects for Postdoctoral Research Funds [1302007A]
  11. Economical Forest Cultivation and Utilization of Collaborative Innovation Center in Hunan Province

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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.

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