Temperature-dependent selective purification of plasmid DNA using magnetic nanoparticles in an RNase-free process

Zhi Shan; Chenglei Li; Xu Zhang; Ken D Oakes; Mark R Servos; Qi Wu; Hui Chen; Xianxiang Wang; Qianming Huang; Yi Zhou; Wanshen Yang

doi:10.1016/j.ab.2011.01.012

Temperature-dependent selective purification of plasmid DNA using magnetic nanoparticles in an RNase-free process

Anal Biochem. 2011 May 1;412(1):117-9. doi: 10.1016/j.ab.2011.01.012. Epub 2011 Jan 15.

Authors

Zhi Shan¹, Chenglei Li, Xu Zhang, Ken D Oakes, Mark R Servos, Qi Wu, Hui Chen, Xianxiang Wang, Qianming Huang, Yi Zhou, Wanshen Yang

Affiliation

¹ Faculty of Science, Sichuan Agricultural University, Yaan 625014, People's Republic of China.

PMID: 21241655
DOI: 10.1016/j.ab.2011.01.012

Abstract

Carboxyl group-functionalized magnetic nanoparticles were used to develop an RNase-free method for plasmid DNA (pDNA) purification directly from RNA-containing crude Escherichia coli lysates. This method takes advantage of differing adsorption behaviors of pDNA and RNA onto magnetic nanoparticle surfaces at different temperatures. Pure pDNA can be isolated between 70 and 80°C without sacrificing DNA quality and quantity, as evidenced by comparison with that obtained using organic solvents or commercial kits. This RNase-free method is rapid, simple, cost-effective, and environmentally friendly, and it can be easily scaled up for the production of pharmacological-grade pDNA.

MeSH terms

2-Propanol / chemistry
Chemical Precipitation
DNA / isolation & purification*
Magnetics*
Nanoparticles / chemistry*
Plasmids / chemistry
Plasmids / isolation & purification*
Ribonucleases / chemistry*
Ribonucleases / isolation & purification
Temperature

Substances

DNA
Ribonucleases
2-Propanol