体积排阻液相色谱法质粒的含量测定

首次以体积排阻液相色谱法进行质粒提取液浓度的测定研究,通过纯质粒溶液在260 nm处的紫外吸峰面积与质粒浓度之间数据的线性最小二乘拟合,得到了反映质粒浓度和紫外吸收峰峰面积之间关系的标准曲线:Y=2×10~(-5) X,通过验证实验,确定了质粒提取液中不同组分,如核糖核酸,蛋白质等在色谱图中的出峰位置.结合标准曲线测定了自制的质粒提取液的质粒浓度为0.32 mg/mL.该方法方便,快捷,准确.

The Determination of Plasmid Concentration in lysate by Size Exclusion Chromatography

This study was focus on the determination of plasmid concentration in the lysate by size exclusion chromatography(SEC).At first,the pure plasmid solution was prepared by Mini preps DNA purification system,then the peak area of the plasmid in the chromatogram was determined with different plasmid concentration.The relationship between the plasmid peak area in the chromatogram and the plamid concentration can be expressed Y = 2×10~(-5) X with linear-least square fitting method.The position in the chromatogram of different content in the lysate was determined by experiments.The plamid concentration of the lysate was get(0.32 mg/mL)with the plasmid peak area and the equation of 7 = 2×10~(-5) X.This method is convenient and accurate.