Plasmid DNA isolation using amino-silica coated magnetic nanoparticles (ASMNPs).
A simple and efficient approach for the rapid isolation of plasmid DNA from crude cell lysates has been described. The approach took advantage of the amino-modified silica coated magnetic nanoparticles (ASMNPs) with positive zeta potential at neutral pH and superparamagnetism under the external magnetic fields. As a demonstration, the pEGFP-N3 plasmid has been concentrated and isolated from the E. coli DH5alpha transformed with pEGFP-N3 plasmid through electrostatic binding between the positive charge of the amino group of ASMNPs and the negative charge of the phosphate groups of the plasmid DNA. Then the pEGFP-N3 plasmid has been released easily and quickly from the pEGFP-N3 plasmid-ASMNPs complexes with 3M NaCl. The entire procedure could be carried out by the aid of external magnetic fields in 15min and eliminate the need of phenol, cesium chloride gradients or other noxious reagents and complexes operation. Moreover, the pEGFP-N3 plasmid obtained by this approach retains biological activity that can be suitable for restriction enzyme digestion and cells transfection with expression of green fluorescence protein.
He
Xiaoxiao
X
State Key Laboratory of Chemo/Biosensing and Chemometrics, Engineering Center for biomedicine, Institute of Life Science & Biological Technology, Key Laboratory for Bio-Nanotechnology and Molecule Engineering of Hunan Province, Changsha 410082, PR China.
Huo
Hailing
H
Wang
Kemin
K
Tan
Weihong
W
Gong
Ping
P
Ge
Jia
J