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化学学报
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化学学报 ›› 2010, Vol. 68 ›› Issue (9): 917-920. 上一篇    下一篇

研究论文

金纳米颗粒介导不对称PCR: 制备单链核酸的新方法

孟祥贤，羊小海，王柯敏*，郭秋平，黄青山   

  1. (湖南大学化学生物传感与计量学国家重点实验室 化学化工学院 生物医学工程中心 生物纳米与分子工程湖南省重点实验室 长沙 410082)
  • 投稿日期:2009-03-06 修回日期:2009-07-26 发布日期:2010-01-10
  • 通讯作者: 孟祥贤 E-mail:xiangxianmeng@yahoo.com.cn

Gold Nanoparticle-Based Asymmetric PCR for Single-Strand DNA

MENG Xiang-Xian, YANG Xiao-Hai, WANG Ke-Min, GUO Qiu-Beng, HUANG Jing-Shan   

  1. (State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Biomedical Engineering Center, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University, Changsha 410082)
  • Received:2009-03-06 Revised:2009-07-26 Published:2010-01-10
  • Contact: 孟 祥贤 E-mail:xiangxianmeng@yahoo.com.cn

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被引次数

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单链核酸杂交技术是现代医学生物学等领域诊断与检测的重要手段. 传统不对称PCR制备单链核酸的方法, 需要严格控制引物和模板数量及反复摸索退火温度, 操作复杂. 发展了一种利用金纳米颗粒介导的不对称PCR快速制备单链核酸的新方法. 浓度为0.1～0.5 nmol/L金纳米颗粒可显著改善PCR扩增特异性, 提高扩增效率. 在不对称PCR扩增中, 加入合适浓度的金纳米颗粒方便快捷得到单链核酸产物, 不需要进行复杂的条件优化等操作步骤. 通过快速制备地中海遗传病两个点突变CD17 (A→T)和 IVS-2-654 (C→T)位点的单链DNA靶序列, 证实该方法是一种简便、有效的获得单链核酸的方法, 有望在单链核酸技术领域发挥重要作用.

关键词: 金纳米颗粒, 不对称PCR, 单链核酸, 地中海贫血病

Hybridization technique for single-strand DNA (ssDNA) is an important strategy in modern biomedical engineering. Researches on ssDNA through asymmetric PCR generally include complex work and are time-consuming. In this paper, a novel approach was developed for asymmetric PCR based on gold nanoparticle. It was found that appropriate concentrations of gold nanoparticles can enhance significantly the efficiency of asymmetric PCR to obtain ssDNA without complex process. Using the proposed method, ssDNA prodcucts of CD17 (A→T) and IVS-2-654 (C→T) sites in the β-thalassemia has been achieved conveniently and rapidly.

Key words: gold nanoparticle, asymmetric PCR, ssDNA, β-thalassemia