一种用于PCR模板制备的电泳产物简易回收方法  

A Simple Way of Agarose Gel DNA Fragments Recovery for PCR Amplification

作　　者：王洪敏[1] 马文丽[1] 黄海[1] 郑文岭[2] 

机构地区：[1]第一军医大学分子生物学研究所,中国广东广州510515 [2]广州军区广州总医院肿瘤分子生物学研究所,中国广东广州510010

出　　处：《生命科学研究》2003年第4期365-368,共4页Life Science Research

基　　金：国家自然科学基金资助项目(39880032);广州市重点科技攻关项目(99 Z 022 01)

摘　　要：为了探索一种简便、有效而且能从琼脂糖凝胶中大量回收用于第2次PCR扩增的DNA电泳条带的方法,采用刀片切胶法和牙签插胶法从琼脂糖中回收DNA,并进行了两种方法的比较.结果显示牙签插胶法回收的DNA用作第2次PCR的模板,获得了清晰、稳定的PCR产物电泳条带,用该法成功地制备了一批DNA微阵列探针.由此可见牙签插胶法是一种简便、快速、有效的用于PCR模板的DNA琼脂糖凝胶回收法.To study a simple, effective method for DNA recovering for subsequent secondary PCR amplification from agarose gel in large scale. Two methods were tested and compared. The first approach is to cut the DNA band with a lancet from the agarose gel after electrophoresis; the second one is to insert briefly the wooden toothpick tip inside the DNA bands. The DNA were eluted and used as PCR template for the secondary PCR re-amplification. Electrophoresis showed clear and consistent bands after PCR re-amplification of the DNA recovered from the agarsose gel by both method, but the toothpick insertion method is much easier and quicker; a number of DNA microarray probes were successfully prepared using this method. It could be concluded that the method of toothpick insertion is an efficient method for secondary PCR template preparation.

关 键 词：PCR模板制备 电泳产物 回收方法 聚合酶链反应 DNA微阵列 探针制备 基因芯片 

分 类 号：Q503[生物学—生物化学] Q52