从富含酚类的茶类植物叶中提取纯净的总DNA

Extracting the Total DNA of High Purity from the Leave of Several Species of Camellia Sect. Thea

采用三％十六烷基三甲基溴化胺（ＣＴＡＢ）方法，改进研磨方法并适当延长研磨时间至２０－３０ｍｉｎ。选择合适的保温温度和保温时间，为６０℃条件下水浴保温１．５ｈ。从野生可可茶（Ｃａｍｅｌｌｉａ ｐｔｉｌｌｏｐｙｌｌａ）和栽培茶类（Ｃ．ａｓｓａｍｉｃａ，Ｃ．ｓｉｓｎｅｎｓｉｓ）植物中提取到量多且较纯的总ＤＮＡ，平均含量为１２６７ｎｇ／μＬ，能直接用于ＰＣＲ扩增。经玻璃粉纯化系统（ＤＰＳ）纯化后，获得高纯度的ＤＮＡ，平均含量约为１３６ｎｇ／μＬ，纯化回收率平均为１１％，能完全满足ＲＡＰＤ、ＡＲＬＰ、ＲＦＬＰ分析及测序等分子生物学实验。

The total DNA were extracted out from five materials of Camellia Sect. Thea by the modified 2 × CFAB (Cetyltrimethyl Ammonium Bromide) method. The both times spent in grinding with liquid nitrogen arid in heating with constant temperature (60℃) were lengthened properly, being about 20 ~ 30 mm and 1.5 h respectively. The DNA of high purity is obtained after purified by Glass Milk (DPS). The average concentration of total DNA is 1 267 ng/μL before purified, and 136 ng/μL after purified. The average returning rate of DNA is 11 %. The total DNA extracted can be applied effectively in PCR, RAPD, AFLP and RFLP analysis, Sequencing and other molecular biological researches.