LI Zhen-qing,CHEN Qin,ZHU Jian-dong,ZENG Yuan,ZHANG Da-wei,Yoshinori Yamaguchi.Research on Quantitation of DNA Based on Gel Electrophoresis Chip[J].Journal of Instrumental Analysis,2025,44(05):931-935.
LI Zhen-qing,CHEN Qin,ZHU Jian-dong,ZENG Yuan,ZHANG Da-wei,Yoshinori Yamaguchi.Research on Quantitation of DNA Based on Gel Electrophoresis Chip[J].Journal of Instrumental Analysis,2025,44(05):931-935. DOI: 10.12452/j.fxcsxb.240911387.
Research on Quantitation of DNA Based on Gel Electrophoresis Chip
为确定凝胶电泳后条带中DNA的长度和浓度,该文提出了一种通过DNA荧光强度精确分析DNA长度及浓度的有效方法。对D2000 DNA ladder进行凝胶电泳,采集图像并进行处理。利用荧光强度与DNA浓度之间的线性关系以及迁移距离与DNA长度的线性关系进行数据分析。结果显示:采用SYBR Green I作为荧光染料电泳DNA时,图像中各像素点中的绿色与红色灰度值可以反映DNA的特征信息,且绿色灰度值明显高于红色灰度值;对于小于2 000 bp的DNA片段,其分子量与迁移距离呈反比例函数关系,当二者分别取对数时,其相关系数达0.971,可用于建立DNA分子量与迁移距离的数学模型以计算DNA分子量;通过计算电泳峰峰值及电泳峰积分面积,发现对于浓度比为2∶1的两个DNA条带,二者的比值为2.16与1.96、2.11与1.94、2.16与1.93,表明电泳峰积分面积能更有效地反映DNA的真实浓度值。该研究对于核酸凝胶电泳仪的开发具有重要应用价值。
Abstract
To determine the DNA length and concentration of the bands after gel electrophoresis,this paper proposes an effective method to accurately analyze the length of DNA and its concentration by DNA fluorescence intensity from slab gel electrophoresis. Perform gel electrophoresis on the D2000 DNA ladder,collect image data,and process the images.The linear relationship between fluorescence intensity and DNA concentration,and the linear relationship between migration distance and DNA length were used for data analysis.Experimental results show that:When SYBR Green I was used as the fluorescent dye for electrophoresis of DNA,the green and red grey values in each pixel could reflect the characteristic information of DNA,and the green grayscale value was obviously higher than the red one. For the DNA fragments less than 2 000 bp,the migration distance were inversely proportional to the molecular weight,especially when the two value was calculated based on logarithmic,their correlation coefficient was 0.971,which could be used to establish a mathematical model for the calculation of DNA molecular weight based on its migration distance. By calculating the peak value and the peak area of the electropherogram,it was found that the ratios of the two DNA bands with the concentration ratio of 2∶1 were 2.16 and 1.96,2.11 and 1.94,2.16 and 1.93,indicating that the electrophoretic peak area was more effective in calculating the concentration value of DNA. This study has important application value for the development of nucleic acid gel electrophoresis instrument.
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