聚合酶链反应产物的HPLC定量分析

HPLC Quantitative Analysis of Polymerase Chain Reaction (PCR) Products

建立了ＨＰＬＣ分析聚合酶链反应（ＰＣＲ）产物的方法，探讨了梯度、流速、柱温等条件对相对分子质量参照物λ－ＤＮＡ－ＨｉｎｄⅢ分离的影响，ＮａＣｌ以４０～５０ｍｍｏｌ／ｍｉｎ的梯度变化，流速为１ｍＬ／ｍｉｎ时分离效果最佳。该法精密度和回收率高、线性良好，可用于ＰＣＲ扩增条件优化时的相对定量分析。

A method for the analysis of the polymerase chain reaction(PCR) products by HPLC was developed. The Effect of conditions, such as gradient of elution, flow rate, column temperature, etc on the separation of λ _ DNA-Hind Ⅲwas examined. The separation was performed on a non _ porous ion exchange column with gradient elution of sodium chloride in 20 mmol/L Tris-HCl buffer (pH 9.0) at a flow rate of 1.0 mL/min, the detection wavelength was at 260 nm. λ _ DNA-Hind Ⅲdigest and β _ actin PCR product were analyzed with high recovery and good linear relationship.