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3.8 试剂加入顺序对体系荧光的影响 固定其它条件，考察试剂加入顺序对BCDO-DNA体系荧光猝灭的影响，结果如图3.21和3.22所示。 根据图3.21和3.22测定的荧光强度作BCDO-DNA体系荧光猝灭强度ΔF与试剂加入顺序的变化曲线。结果如图3.23所示。本实验中试剂的不同的加入顺序对BCDO与DNA体系的影响都很大。但缓冲溶液在最后加入时，BCDO-DNA体系的荧光猝灭值ΔF最大。因此采用缓冲溶液最后加入。 由以上实验条件的选择得到测定BCDO-DNA体系的较佳实验条件，DNA的浓度为5.0mg•L-1，BCDO的浓度为3.0×10-5mol•L-1，pH=6.8时，反应时间为5min，溶剂体积小于10%，溶液温度为室温，加入顺序为BCDO+缓冲溶液+DNA。根据较佳实验条件，对BCDO-DNA体系的干扰试验，工作曲线，作用方式，进行了研究，并通过EB法作对照，对BCDO-DNA体系的猝灭方式进行了进一步的判断。 |
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3.8 The effect of the addition order of reagents on the fluorescence of the system Other factors were fixed, the effect of the addition order of reagents on the fluorescence quenching of the BCDO-DNA system were investigated, the results were shown in Figure 3.21 and 3.22. According to the fluorescence intensity investigated in Figure 3.21 and 3.22, the curve was made of the strength of fluorescence quenching (ΔF) of the BCDO-DNA system and the addition order of reagents, as was shown in Figure 3.23. In this experiment, the different addition order of reagents had a great influence on the system both BCDO and DNA. However, when the buffer solution was added at last, the maximum fluorescence quenching value of the BCDO-DNA system was obtained. So, the buffer solution should be added finally. From the selection of above experiment condition, the better experimental factors determining the BCDO-DNA system were made, the concentration of DNA and BCDO were 5.0mg•L-1 and 3.0×10-5mol•L-1, respectively; pH was 6.8, the reaction time was 5min, the volume of solvent was less than 10%, the temperature of the solution was room temperature(20℃), the addition order was BCDO + buffer solution + DNA. According to the better experimental condition, studies of the interference test, working curve, action mode of the BCDO-DNA system were made, compared with the EB method, the quenching method of the BCDO-DNA system was further determined. 翻译的不好，呵呵，抛砖引玉哈~~~ |
2楼2012-04-27 14:56:38