地高辛3prime;末端标记检测效率的分析.pdfVIP

安徽农业科学,Journal of Anhui Agri . Sci .2008 ,36( 12) :4888 - 4889 ,4893 责任编辑 罗芸 责任校对 李洪 地高辛3′末端标记检测效率的分析 郑海霞, 王玉国, 冯智富 ( 山西农业大学农学院, 山西太谷 030801) 摘要 [ 目的] 为核酸杂交试验提供依据。[ 方法] 利用3 个不 同长度的核酸探针 进行3′末端地 高辛标记, 并对标 记效率进行检测, 研 究 探针 的长度和浓度及其在尼龙膜上的固定方法对其信号强度的影响。[ 结 果] 聚 丙烯酰胺凝胶 电泳结果表 明地 高辛标记的探针 比未标 记探针滞后, 说明标记较为成功。检测信号强度随探针摩 尔浓度的降低 而降低 。探针的固定方法对信号强度也有一定的影响。紫外交 联和 高温烘烤均能使检测信号增强, 而使 用紫外交联的信号强度较 高。[ 结论] 地高辛检测的信号强度与探针 的长度、浓度、固定方法以 及核酸与膜 的结合效率有关。 关键词 核酸探针; 地 高辛标记; 标记效率 中图分类号 S188 文献标识码 A 文章编号 0517 - 6611(2008) 12 - 04888 - 02 Analysis of the Detection Efficiency of Digoxin Labeling at 3′End ZHENG Hai-xia et al ( College of Agronomy , Shanxi Agricultural University , Taigu ,Shanxi 030801) Abstract [ Obj ective] The ai m of the research was to provide basis for nucleic acid hybridization test . [ Method] Three probes of nucleic acid with dif- ferent length were used to label at 3′end with Digoxin and detect its labeling efficiency . The effects of the length and concenstration of probes and its fixa- tion methods on nylon membrane on the signal intensity were studied . [ Result ] The results of polyacrylamide gel electrophoresis showed that the probes with Digoxin labeling was hysteresis than that without labeling , which indicated that the labeling was more successful . The detected signal intensity was re- duced with the reduction of concenstration of probe .The fixation method of probe also had certai n effects on the signal intensity .Both ultraviolet cross-li nk- ing and high temperature firing coul d make the detection signal be strengthened . And the signal intensity at ultraviolet cross- linking was higher . [ Conclu- sion] The detected signal i ntensity of Digoxin was related withthe length ,concentration ,fixation and the co mbined efficacy between nuceleic acid and me m- brane . Key words Nucleic acid probe ;Digoxi n label