生物技术通报Issue(12)：105-109,5.DOI:10.13560/j.cnki.biotech.bull.1985.2015.12.015
转基因甘蔗植株Southern杂交体系的优化
The Optimization of Southern Blot for Transgenic Sugarcane Plants
摘要
Abstract
Using sugarcane as materials, the DIG-labeled Southern blot was optimized through several key aspects:the comparison of different-labeled probe methods, extraction of sugarcane genome DNA, the amount of enzyme digestion of genome DNA, enzyme digestion time and a serial of procedures in the process of the hybrid. The results showed that sugarcane DNA extracted by the improved CTAB method met the requirements of the latter experiments, the efficiency of PCR-labeled probe was higher than that of random primer-labeled probe, so PCR-labeled probe method was suitable for the Southern blot, 40μg DNA samples in enzyme digestion system of 400μL for 10 hours could achieved desirable digestion result;while hybrid temperature was 40℃ and hybridizing time was 18 h, a clear hybridization band could be observed. The optimization of sugarcane Southern blot provides a reference for the analysis of Southern blot of transgenic sugarcane.关键词
甘蔗/Southern杂交/地高辛/PCR法标记探针/酶切Key words
sugarcane/Southern blot/digoxigenin/PCR-labeled probe/enzyme digestion引用本文复制引用
崔学强,张树珍,沈林波,冯翠莲..转基因甘蔗植株Southern杂交体系的优化[J].生物技术通报,2015,(12):105-109,5.基金项目
国家自然科学基金项目（31101197），现代农业产业技术体系建设专项基金 （）
