Position: Home > Articles > A Method of Improved Southern Blot Hybrid
Xinjiang Agricultural Sciences
2007,44
(3)
116-118
一种改良的Southern印迹杂交方法
作 者：
李建平;张燕红;王冬梅;黄全生
单 位：
新疆农科院核生所
关键词：
Southern印迹;DNA;杂交;植物基因组
摘 要：
在综合前人方法的基础上,改良了一种简捷、灵敏、廉价、效果可靠的Southern印迹杂交方案,使用0.4mol/L NaOH溶液将琼脂糖凝胶上的DNA在变性的同时转移到尼龙膜上,紫外交联固定后用于杂交。预杂交液和杂交液选用Church缓冲系统,该缓冲系统中仅需10 g/L BSA作为封闭剂即可。整个洗膜过程可简化为一种溶液、2~3次洗涤。碱变性及转移后的尼龙膜,也可以用于地高辛(DIG)等非放射性检测系统,并可耐多轮杂交和洗膜。方法可适用于包括高等植物基因组在内的多种生物材料的DNA检测。
译 名：
A Method of Improved Southern Blot Hybrid
作 者：
LI Jian-ping,ZHANG Yan-hong,WANG Dong-mei,HUANG Quan-sheng(Institute of Nuclear and Biological Technology,Xinjiang Academy of Agricultural Sciences,Urumqi 830091,China)
关键词：
Southern blot;DNA;hybridization;plant genome
摘 要：
A simple,rapid,sensitive,low-cost and reproducible protocol of Southern blot analysis was developed based on a comprehensive analysis of the previous reports.With a solution of 0.4mol/L NaOH,DNA in the agarose gel can be denatured and transferred onto the nylon membrane in a single step.The Church buffer system is the only buffer which is used as pre-hybridization and hybridization.The whole process of washing the membrane can be simplified as one solution.The prepared membrane can also be used in the non-radioactive DNA detection system.This method is applied to detect target DNA such as plants which hold comparatively large genome.
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