Attaching thiolated DNA on gold nanoparticles(AuNPs)is a milestone of nanobiotechnology,which combines the programmability and molecule recognition properties of biopolymers with the optical,thermal and catalytic properties of inorganic nanomaterials.Current standard protocols to attach thiolated DNA on AuNPs involve multiples steps and are time-consuming and labor-intensive,which greatly restrict their applications.Here we demonstrate a rapid and facile strategy to attach thiolated DNA on AuNPs based on the excellent stabilization effect of mPEG-SH.AuNPs with a wide range of size are first protected by mPEG-SH in the presence of Tween 20,resulting in excellent stability in high ionic strength and extreme pH.High concentration of NaCl can be directly added to the mixture of DNA and AuNP,which allows highly efficient DNA attachment to AuNP surface by minimizing the electrostatic repulsion.The whole DNA loading processes can be simply completed in 2 hr.DNA loaded on AuNPs are stable for half a month and yet precisely hybridize with the complementary sequences,which can be applied to prepare core-satellite nanostructure.Moreover,cytotoxicity assay confirms that the DNA-AuNPs synthesized by this method bare lower cytotoxicity than that by current standard methods.The proposed method provides a new way to stabilize AuNPs for fast and facile loading thiolated DNA on AuNPs and will find wide applications in many areas requiring DNA-AuNPs.