快速小量提取小麦叶片DNA的一种简易方法

目的]寻求少量、快速提取小麦幼叶DNA的方法。方法]以小麦幼叶（只要是健康的绿叶，老叶也可）为试验材料，用改良的CTAB法小量快速提取转基因小麦总DNA。具体步骤为：①直接用已灭菌的1．5ml离心管夹取小麦叶片（无需称量），然后在液氮中快速冷冻后用玻璃棒研磨成细粉（研磨直接在离心管中进行，不必先在研钵中研磨然后再转移到离心管里，能够有效减少材料损失），加入600山预热（65℃）的2％CTAB提取缓冲液，快速振荡摇匀。置于65℃的水浴锅中温育30min，期间温和混匀几次。②取出离心管，冷却至室温，加入600m氯仿：异戊醇（24：1）充分混匀，于4℃下12000r／min离心8min（只抽提1次）。③取上清，加入等体积预冷的异丙醇，缓慢混匀，至有絮状DNA析出，-20℃放置30min以上。④4℃下12000r／min离心8min。弃上清液，将沉淀用700μl70％乙醇清洗2次，离心，弃乙醇，室温下挥发乙醇，加入适量ddH：O或0．1×TE溶解沉淀（DNA），-20℃保存。所提取的DNA以0．8％琼脂糖凝胶电泳检测。结果]改良的CTAB法提取小麦基因组DNA所提取的DNA纯度高、无降解现象，适用于进行常规PCR扩增，并且需要材料量少、成本低，还可缩短操作周期、节省时间。结论]该研究为小麦DNA的提取提供了一种所需材料量少、简便、快速的方法。

A Simple and Quick Method of Extracting Genomic DNA from Wheat Leaves

Objective] The aim was to seek a simple and quick method of extracting genomic DNA from wheat leaves. Method] Taking tender leaves of wheat as test materials, total DNA of transgenic wheat was extracted by using modified CTAB method. The extracted DNA was detected by 0.8% agarose gel electrophoresis. Result] DNA purity of extracted genome DNA from wheat was high and no degradation phenomenon using modified CTAB method, and was suitable for carrying out normal PCR amplification. Conclusion] This study provides a simple and quick method for extracting DNA from wheat with a spot of material.