Fluorescent Quantification of DNA Based on Core-Shell Fe3O4@SiO2@Au Nanocomposites and Multiplex Ligation-Dependent Probe Amplification.
In this research, a novel method for relative fluorescent quantification of DNA based on Fe3O4@SiO2@Au gold-coated magnetic nanocomposites (GMNPs) and multiplex ligation- dependent probe amplification (MLPA) has been developed. With the help of self-assembly, seed-mediated growth and chemical reduction method, core-shell Fe3O4@SiO2@Au GMNPs were synthesized. Through modified streptavidin on the GMNPs surface, we obtained a bead chip which can capture the biotinylated probes. Then we designed MLPA probes which were tagged with biotin or Cy3 and target DNA on the basis of human APP gene sequence. The products from the thermostable DNA ligase induced ligation reactions and PCR amplifications were incubated with SA-GMNPs. After washing, magnetic separation, spotting, the fluorescent scanning results showed our method can be used for the relative quantitative analysis of the target DNA in the concentration range of 03004~0.5 µM.
Jing
J
Fan
Haowen
H
Yang
Ming
M
Liu
Dan
D
Wu
Hongrong
H
Jiang
Xin
X
Zeng
Sauli
S
Elingarami
Zhiyang
Z
Ll
Song
S
Li
Hongna
H
Liu
Nongyue
N
He
D001483:Base Sequence; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods; D004247:DNA / Q000032:analysis* / Q000235:genetics*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006046:Gold / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods; D008969:Molecular Sequence Data; D060885:Multiplex Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods; D053761:Nanocomposites / Q000737:chemistry / Q000648:ultrastructure*; D017422:Sequence Analysis, DNA / Q000295:instrumentation / Q000379:methods; D012822:Silicon Dioxide / Q000737:chemistry
D006046:Gold; D012822:Silicon Dioxide; D004247:DNA