一种采用荧光猝灭法对核酸进行灵敏测定的方法。
The sensitive determination of nucleic acids using a fluorescence-quenching method.
作者信息
Jia Zhen, Yang Jinghe, Wu Xia, Sun Changxia, Wang Fei, Liu Shufang, Wang Feng
机构信息
Key Laboratory of Colloid and Interface Chemistry of the Education Ministry, School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, People's Republic of China.
出版信息
Luminescence. 2006 Jul-Aug;21(4):207-13. doi: 10.1002/bio.906.
Experiments indicated that nucleic acids can quench the fluorescence of the Eu3+ -2-thenoyltrifluoroacetone (TTA)-1,10-phenanthroline (Phen) system. Based on this, a sensitive method for the determination of nucleic acids was proposed. The experiments indicated that under the optimum conditions, the quenched fluorescence intensity was in proportion to the concentration of nucleic acids in the range 1.0 x 10(-11)-1.0 x 10(-6) g/mL for yeast RNA (yRNA), 5.0 x 10(-11)-5.0 x 10(-7) g/mL for fish sperm (fsDNA) and 1.0 x 10(-10)-1.5 x 10(-6) g/mL for calf thymus DNA (ctDNA). Their detection limits were 3.0 x 10(-12), 4.0 x 10(-12) and 5.0 x 10(-11) g/mL, respectively. Therefore, the proposed method is one of the most sensitive methods available. The interaction between nucleic acids and Eu3+ -TTA-Phen is also discussed.
实验表明，核酸能够猝灭铕（Eu3+）-噻吩甲酰三氟丙酮（TTA）-1,10-菲啰啉（Phen）体系的荧光。基于此，提出了一种测定核酸的灵敏方法。实验表明，在最佳条件下，猝灭后的荧光强度与核酸浓度成正比，对于酵母RNA（yRNA），浓度范围为1.0×10−11 - 1.0×10−6 g/mL；对于鱼精DNA（fsDNA），浓度范围为5.0×10−11 - 5.0×10−7 g/mL；对于小牛胸腺DNA（ctDNA），浓度范围为1.0×10−10 - 1.5×10−6 g/mL。它们的检测限分别为3.0×10−12、4.0×10−12和5.0×10−11 g/mL。因此，所提出的方法是现有最灵敏的方法之一。还讨论了核酸与Eu3+ -TTA-Phen之间的相互作用。
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