Temperature-dependent selective purification of plasmid DNA using magnetic nanoparticles in an RNase-free process.
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.
Faculty of Science, Sichuan Agricultural University, Yaan 625014, People's Republic of China.
Zhi
Z
Shan
Chenglei
C
Li
Xu
X
Zhang
Ken D
KD
Oakes
Mark R
MR
Servos
Qi
Q
Wu
Hui
H
Chen
Xianxiang
X
Wang
Qianming
Q
Huang
Yi
Y
Zhou
Wanshen
W
Yang
D019840:2-Propanol / Q000737:chemistry; D011232:Chemical Precipitation; D004247:DNA / Q000302:isolation & purification*; D008280:Magnetics; D053758:Nanoparticles / Q000737:chemistry*; D010957:Plasmids / Q000737:chemistry / Q000302:isolation & purification*; D012260:Ribonucleases / Q000737:chemistry* / Q000302:isolation & purification; D013696:Temperature
D004247:DNA; D012260:Ribonucleases; D019840:2-Propanol