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

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Abstract

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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Authors and Affiliations

  1. State Key Laboratory of Bioelectronics; School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096, China

    Jiuhai Wang, Zeeshan Ali, Hongna Liu, Fu Li, Haowen Yang, Lei He & Nongyue He

  2. Clinical Laboratory, the Second Hospital of Nanjing, Nanjing, 210003, China

    Nianyue Wang

  3. Jiangsu Province Blood Center, Nanjing, 210042, China

    Wenbiao Liang

  4. Economical Forest Cultivation and Utilization of 2011 Collaborative Innovation Center in Hunan Province; Hunan Key Laboratory of Green Packaging and Application of Biological Nanotechnology, Hunan University of Technology, Zhuzhou, 412007, China

    Libo Nie & Nongyue He

  5. Department of Laboratory Medicine, the Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210011, China

    Zhiyang Li

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  1. Jiuhai Wang
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  2. Zeeshan Ali
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  3. Nianyue Wang
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  4. Wenbiao Liang
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  6. Fu Li
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  8. Lei He
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  9. Libo Nie
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  10. Nongyue He
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  11. Zhiyang Li
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Correspondence to Nongyue He or Zhiyang Li.

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Wang, J., Ali, Z., Wang, N. et al. Simultaneous extraction of DNA and RNA from Escherichia coli BL 21 based on silica-coated magnetic nanoparticles. Sci. China Chem. 58, 1774–1778 (2015). https://doi.org/10.1007/s11426-015-5483-x

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  • DOI: https://doi.org/10.1007/s11426-015-5483-x

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