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[1] Nie Libo, Chen Hong, Tan Meijun, et al. Improvement of hybridization signalsof gold label silver stain gene detection [J]. Journal of Southeast University (English Edition), 2004, 20 (4): 463-466. [doi:10.3969/j.issn.1003-7985.2004.04.014]
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Improvement of hybridization signalsof gold label silver stain gene detection()
金标银染基因检测方法中杂交信号的提高
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Journal of Southeast University (English Edition)[ISSN:1003-7985/CN:32-1325/N]

Volumn:
20
Issue:
2004 4
Page:
463-466
Research Field:
Chemistry and Chemical Engineering
Publishing date:
2004-12-30

Info

Title:
Improvement of hybridization signalsof gold label silver stain gene detection
金标银染基因检测方法中杂交信号的提高
Author(s):
Nie Libo1;  Chen Hong2;  3;  Tan Meijun3;  He Nongyue1
1Key Laboratory of Molecular and Biomolecular Electronics of Ministry of Education, Southeast University, Nanjing 210096, China
2Department of Life Science and Technology, Central South Forestry University, Changsha 410004, China
3School of Packaging and Printing, Zhuzhou Institute of Technology, Zhuzhou 412008, China
聂立波1;  陈洪2;  3;  谭美君3;  何农跃1
1东南大学分子与生物分子电子学教育部重点实验室, 南京 210096; 2中南林学院生命科学技术系, 长沙 410004; 3株洲工学院包装印刷学院, 株洲 412008
Keywords:
gold label silver stain;  gold nanoparticle;  hybridization signal
金标银染;  纳米金;  杂交信号
PACS:
O633
DOI:
10.3969/j.issn.1003-7985.2004.04.014
Abstract:
The factors that influence the colorimetric gene detection of gold label silver stain and improve the detection signals are studied. The influence of amino DNA probes and thiol DNA modified gold nanoparticles is investigated based on a sandwich hybridization system. An increase in amino probe concentration brings about an increase in hybridization signal which reaches a threshold corresponding to the saturated concentration of amino probes bounded onto a glass slide surface. Since the steric hindrance effect of nanoparticles is dominant over the influence of a surface area, the bigger gold nanoparticles lead to weaker hybridization signals. The hybridization efficiency enhances significantly with the increase of the thiol DNA modified nanoparticle concentrations. Experimental results show that 125 μmol/L of the amino DNA probe concentration, 15 nm of the gold nanoparticle diameter, and 4. 07 nmol/L of the thiol DNA modified gold nanoparticle concentration are optimal for the detection system. The hybridization signals can be improved remarkably by choosing optimal hybridization conditions.
研究了金标银染基因检测方法中的影响因素以及提高DNA杂交信号的途径. 在“三明治”杂交系统中, 着重研究了氨基DNA探针和纳米金标记巯基DNA探针对杂交的影响. 随着氨基探针浓度的增加, 杂交信号增加, 探针达到一定浓度后信号不再增加, 形成一个平台, 说明连接于玻片表面的探针已达到饱和. 由于纳米金颗粒空间位阻效应的影响大于纳米颗粒表面积的影响, 标记的纳米金颗粒越大, 杂交信号越弱. 而随着金标巯基DNA探针浓度的增加, 杂交效率显著增加, 导致杂交信号提高. 优化后的杂交条件为: 氨基DNA探针浓度为125 μmol/L, 纳米金粒径为15 nm, DNA标记后的纳米金颗粒浓度为4. 07 nmol/L. 通过上述杂交条件优化, 可提高杂交银染的信号.

References:

[1] Storhoff J J, Elghanian R, Mucic R C, et al. One-pot colorimetric differentiation of polynucleotides with single base imperfections using gold nanoparticle probes [J]. J Am Chem Soc, 1998, 120(9): 1959-1964.
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Memo

Memo:
Biographies: Nie Libo(1973—), female, graduate; He Nongyue(corresponding author), male, doctor, professor, nyhe1958@163.com.
Last Update: 2004-12-20