测定小牛胸腺DNA含量的荧光新方法研究

采用荧光光谱法对多种荧光试剂与小牛胸腺DNA(ctDNA)结合后的光谱特征做了研究,实验结果发现ctDNA不仅可以使夜蓝(NB)的荧光增强,而且在一定浓度范围之内体系的荧光增强程度与ctDNA的量呈线性关系,据此,建立了一种测定ctDNA含量的荧光新方法。当pH值为6.4,反应温度为25℃,反应时间为30min,NB的浓度为2.0×10-5mol.L-1时,ctDNA的浓度在0.01～0.8 g.mL-1范围内,体系的ΔF与ctDNA的浓度呈良好的线性关系,线性方程为:ΔF=627.85c+6.2547(c:g.mL-1),r=0.9986,检测限为4.2×10-5g.mL-1。本方法首次成功地应用于果树榛子中DNA的测定,回收率为96.58%～102.35%,结果令人满意。

Study on a new fluorescence system for detemination of calf thymus DNA.

The characteristics of spectra, combined ctDNA with the diff gents, have been studied by the method of fluorescence spectroscopy. The e that ctDNA could strengthen significantly the fluorescence intensity of NB. erent kings of fluorescent rea xperimental results discovered With increasing of density of ctDNA,fluorescence intensity of the system of NB-ctDNA got stronger and stronger. Moreover, the rela- tionship between the fluorescence intensity and the amount of ctDNA was linear within certain range. Based on this finding, a new fluorscence probe of ctDNA was discovered and a new fluorescence system for determination of the amount of ctDNA was established. When pH=6.4, the temperature was 25℃, time range was 30mins, the concentration of NB was 2.0 ×10^-5 mol . L- 1, and ctDNA concentration was 0.01 -0.8 g . mL-1 ,the linear relationship was significant. And the linear square was AF=627.85cq-6. 2547 （c： g-.mL-1）, the related coefficient was r =0. 9986,and the detection limit was 4.2 × 10^-5g . mL-1 The proposed method had been successfully applied to the determination of DNA withdrawn from the hazelnuts, and the recoveries were in the range of 96.58% to 102.35% with satisfactory results.