非巯基核酸-纳米金偶联物的制备方法及其应用
首发时间:2015-01-21
摘要:传统的核酸-纳米金偶联物制备基于Au-S共价键实现巯基化寡核苷酸在金纳米粒子表面的自组装。虽然这种方法已经证明在很多应用中有效,但巯基化的核酸序列成本较高,且制备过程繁琐、检测动态范围较窄。连续多个连续碱基A(腺嘌呤)序列对纳米金具有高亲和力,与Au-S化学偶联相比能优先吸附在纳米金的表面。在本研究中,我们发现采用在低pH条件下将多聚腺嘌呤末端核酸(PolyA-DNA)快速吸附到纳米金表面与逐步添加NaCl的方法相结合可以提高PolyA-DNA-AuNPs偶联物的稳定性,可用于DNA的检测。对DNA检测的线性范围为0.3-300 nmol/L,灵敏度可达到0.1 nmol/L。该方法制备的核酸-纳米金具有重复性好、灵敏度高、成本低等优点,适于推广应用。
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The Preparation of Non-thiolated DNA/ Gold Nanoparticles and Its Applications
Abstract:Conjugates of DNA and gold nanoparticles (AuNPs) typically are based on the strong Au-S chemistry to self-assemble thiolated oligonucleotides at AuNPs. While this widely employed strategy has proven highly useful in many applications, but thiolated DNA is high cost and this preparation method is complicated as well as the detection dynamic range is narrow. Poly adenine (polyA) sequences containing multiple consecutive adenines preferentially adsorb Au with high a?nity, even comparable to Au-S chemistry. In the study, we found that combining it that under low pH conditions the PolyA-DNA will be rapidly adsorbed onto the surface of gold nanoparticles with the way of gradual addition of NaCl can improve the stability of PolyA-DNA-AuNPs conjugate.This method can be applied to the detection of DNA with a linear range of 0.3-300 nmol/L and a sensitivity as low as 0.1 nmol/L. This well repeatable, highly sensitive and low cost preparation method of DNA-AuNPs conjugates is suitable for popularization and application.
Keywords: Non-thiol nucleic acid Gold nanoparticle Conjugates DNA Detection
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