使用刚果红共振光散射技术测定纳克级核酸。

Determination of nucleic acids at nanogram level using resonance light scattering technique with Congo Red.

作者信息

Wu Xia, Wang Yuebo, Wang Minqin, Sun Shuna, Yang Jinghe, Luan Yuxia

机构信息

Key Laboratory for Colloid and Interface Chemistry of Education Ministry, School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, Shandong, PR China.

出版信息

Spectrochim Acta A Mol Biomol Spectrosc. 2005 Jan 14;61(3):361-6. doi: 10.1016/j.saa.2004.04.009.

DOI:10.1016/j.saa.2004.04.009

PMID:15582801

Abstract

Based on the enhancement of the resonance light scattering (RLS) of Congo Red (CR) by nucleic acid, a new quantitative method for nucleic acid is developed. In the Tris-HCl buffer (pH 10.5), the weak light scattering of CR is greatly enhanced by addition of nucleic acid and CTMAB, the maximum peak is at 560 nm and the enhanced intensity of RLS is in proportion to the concentration of nucleic acid. The linear range is 1.0 x 10(-9) to 1.0 x 10(-6) g ml(-1), 7.5 x 10(-8) to 1.0 x 10(-6) g ml(-1) and 7.5 x 10(-8) to 2.5 x 10(-6) g ml(-1) for herring sperm DNA, calf thymus DNA and yeast RNA, and the detection limits are 0.019, 0.89 and 1.2 ng ml(-1) (S/N = 3), respectively. Actual biological samples were satisfactorily determined.

摘要

基于核酸对刚果红（CR）共振光散射（RLS）的增强作用，建立了一种新的核酸定量方法。在Tris-HCl缓冲液（pH 10.5）中，加入核酸和十六烷基三甲基溴化铵（CTMAB）后，CR的微弱光散射大大增强，最大峰值位于560 nm处，RLS增强强度与核酸浓度成正比。鲱鱼精DNA、小牛胸腺DNA和酵母RNA的线性范围分别为1.0×10⁻⁹至1.0×10⁻⁶ g ml⁻¹、7.5×10⁻⁸至1.0×10⁻⁶ g ml⁻¹和7.5×10⁻⁸至2.5×10⁻⁶ g ml⁻¹，检测限分别为0.019、0.89和1.2 ng ml⁻¹（S/N = 3）。实际生物样品的测定结果令人满意。