改进TRIZOL法提取基因组DNA

Modified Method for Simultaneous Isolation of Genome DNA with TRIZOL Reagent

目的建立一种在提取总RNA的同时又能制备基因组DNA的方法。方法利用TR IZOL法收集水相沉淀总RNA后余下的中间层和有机相,合并去除蛋白质,再提取基因组DNA。通过紫外分光光度法和电泳法予以鉴定,并以之为模板进行PCR扩增鉴定。结果提取的基因组DNA纯度和浓度俱佳,用于PCR扩增效果良好。结论改进的TR IZOL法可以更加充分地利用实验材料,且可靠易行。

Objective To establish a method of genome DNA isolation with TRIZOL reagent which allows both the extraction of total RNA and preparation of genome DNA at the same time. Methods After the initial treatment with TRIZOL and chloroform, the RNA in the aqueous phase was removed, proteins eliminated and then genome DNA was precipitated with ethanol. Spectrophotometric measurements, electrophoresis and PCR were performed to evaluate the quality and quantity of the DNA. Results The isolated DNA ：was free of RNA.and proteins and had a 260/280 ratio 〉 1.7, and the yield was 6-10 μg/106 cultured cells. The DNA was used for PCR and yielded a better result than that isolated with traditional method. Conclusions The modified method for simuhaneous isolation of genome DNA with TRIZOL reagent was very useful and reliable.