几种微量DNA定量测定方法的比较

A COMPARISION OF SEVERAL METHODS FOR MEASURING TRACE DNA

本研究通过对溴乙淀染色、紫外分光光度计和高效液相色谱(HPLC)等方法的比较,找出了适用于不同情况下,微量DNA的最佳测定方法:1.紫外分光光度计,能准确测量纯净的、浓度在1μg/mL以上的DNA浓度;2.在有标准DNA的情况下,溴乙淀染色法能粗略区别1～5ngDNA;3.HPLC能够快速、准确检测0.21ng以上的DNA,此法简便、灵敏、不受杂质干扰,且能同时对多种微量DNA进行分离和定量。

To measure the amount of DNA, the methods of spectrophotometric determination, ethidium bromide fluorescent quantitation and HPLC are studied in this paper. The results show. 1. Spectrophotometric measurement of the amount of ultraviolet irradiation absorbed by the bases of DNA is simple and accurate, but the sample DNA must be pure enough and its concentration must be higher than lug/mL;2. If the DNA is stained by EtBr,as little as l-5ng of DNA can be estimated by the ultraviolet-induced fluorescent emitted by EtBr (observed by eyes or photography). 3. HPLC can measure as little as 0. 2Ing DNA. Its accurate,sensitive and fast. It can separate and measure several trace DNA molecules simultaneously,,