DNA/RNA-gold nanoparticle (DNA/RNA-AuNP) nanoprobes have been widely employed for nanobiotechnology applications. Here we discovered that both thiolated and non-thiolated DNA/RNA can be efficiently attached to AuNPs to achieve high-stable spherical nucleic acid (SNA) within minutes under a domestic microwave (MW)-assisted heating-dry circumstance. Further studies showed that for non-thiolated DNA/RNA the conjugation is poly (T/U) tag dependent. Spectroscopy, test strip hybridization, and loading counting experiments indicate that low-affinity poly (T/U) tag mediates the formation of a standing-up conformation, which is distributed in the outer layer of such a SNA structure. In further applications study, CRISPR/Cas9-sgRNA (135 bp), RNA from Nucleocapsid (N) gene of SARS-CoV-2 (1279 bp), and rolling circle amplification (RCA) DNA products (over 1000 bp) could be successfully attached on AuNPs, which overcomes the routine methods in long-chain nucleic acid-AuNP conjugation, exhibiting great promise in novel biosensing and nucleic acids delivery strategy. This novel heating-dry strategy has improved the traditional DNA/RNA-AuNP conjugation methods in simplicity, rapidity, cost, and universality.

中文翻译：

---

基于超快、一步和微波加热的 DNA/RNA-AuNP 偶联物合成

DNA/RNA-金纳米颗粒 (DNA/RNA-AuNP) 纳米探针已广泛用于纳米生物技术应用。在这里，我们发现在家用微波 (MW) 辅助加热干燥环境下，硫醇化和非硫醇化 DNA/RNA 都可以有效地连接到 AuNPs 上，以在几分钟内实现高稳定性的球形核酸 (SNA)。进一步的研究表明，对于非硫醇化 DNA/RNA，缀合是多聚 (T/U) 标签依赖性的。光谱学、测试条杂交和加载计数实验表明，低亲和力聚 (T/U) 标签介导了直立构象的形成，该构象分布在这种 SNA 结构的外层。在进一步的应用研究中，CRISPR/Cas9-sgRNA (135 bp)、来自 SARS-CoV-2 核衣壳 (N) 基因的 RNA (1279 bp)、和滚环扩增（RCA）DNA产物（超过1000 bp）可以成功地附着在AuNPs上，克服了长链核酸-AuNP偶联的常规方法，在新的生物传感和核酸递送策略中显示出巨大的前景。这种新颖的加热干燥策略在简单、快速、成本和通用性方面改进了传统的 DNA/RNA-AuNP 偶联方法。