从凝胶中回收银染DNA条带的研究  

Study on the Silver-staining DNA bands recovered from polyacrylamide gels

作　　者：肖学珊[1] 张清炯[1] 黎仕强[1] 贾小云[1] 郭向明[1] 邝志和[1] 申惶煊[1] 

机构地区：[1]中山医科大学中山眼科中心暨卫生部眼科学实验室眼遗传分子生物学实验室,广州510060

出　　处：《临床检验杂志》2002年第1期24-27,共4页Chinese Journal of Clinical Laboratory Science

基　　金：广东省自然科学基金 ;( 960 14 9) ;卫生部优秀青年人才基金 ( 970 16)

摘　　要：目的　比较常规克隆测序与从凝胶中回收银染DNA条带进行直接测序和克隆测序的特点。方法　用单链构像多态法分离杂合性突变DNA样品 ,然后从凝胶中回收变异的银染DNA条带进行直接测序和克隆测序 ,并与常规克隆测序进行比较。结果　从凝胶中回收DNA片段直接测序的正确率超过 97% ,回收片段克隆测序的正确率与常规克隆测序相同 ,达10 0 %。结论　从凝胶中回收DNA片段直接测序和克隆测序较常规克隆测序简便实用 ,在基因克隆、突变鉴定等工作中有一定实用价值。Objective To compare the feasibility,reliability and accuracy of cloning sequencing and direct sequencing for DNA fragments recovered from polyacrylamide gels after silver staining with that of routine cloning sequencing.Methods Heterozygous mutant DNA sample was separated by using single strand conformational polymorphism(SSCP)analysis.The mutant DNA fragments,recovered from polyacrylamide gels after silver staining,were analyzed by cloning sequencing and direct sequencing.The DNA sequence was compared with that from routine cloning sequencing.Results The correctness rate of direct sequencing for recovered DNA is above 97% and that of cloning sequencing is 100% which is the same as that of routine cloning sequencing.Conclusion Sequencing of specific DNA fragments recovered from gels is a rapid,practical and reliable way and is of usefulness in gene cloning and mutational identification.

关 键 词：聚丙烯酰胺凝胶电泳 序列分析 基因突变 基因克隆 脱氧核糖核酸回收 

分 类 号：Q523.8[生物学—生物化学] Q785