杉木DNA提取方法的研究

A Simple and Modified Procedure to Isolate and Characterize Total DNA from Leaves of Cunninghamia lanceolata

在杉木新鲜针叶和干叶ＤＮＡ 提取过程中， 应用ｐＨ 值较低， 无机盐浓度较高的提取缓冲液可沉淀蛋白质， 其中加入２％ 的β－ 巯基乙醇可有效地防止酚类物质使ＤＮＡ 变色． 提取出的ＤＮＡ 样品产率在６０～２００ ｎｇ／ｍ ｇ·ＦＷ， ＤＮＡ 质量和纯度较高， ２６０ｎｍ ／２８０ｎｍ 光密度比值在１８～１９ 之间．所得ＤＮＡ 可直接用于限制性内切酶酶切，并可用于随机引物ＰＣＲ 扩增．

A low pH extraction medium with high salts, which avoids ionization and oxidation of phenolic compounds during tissue grinding and precipitation of large amounts of materials, were successfully used to extract total DNA from both fresh and dry materials of Cunninghamia lanceolata . The DNA yields, quality and purity were characterized. These extracted DNA could be used directly for RAPD analysis, which are useful as molecular genetic markers, and digested with restriction enzymes. A fast, inexpensive and reliable procedure has been developed for detecting the genetic diversity of Cunninghamia lanceolata .