牛4-细胞期胚胎mRNA的RT-PCR检测

凝胶银染法快速检测RT-PCR产物,有利于研究附植前胚胎的基因.选择体外培养的牛单个附植前胚胎,以锚定引物和随机引物RT-PCR扩增,用不同浓度聚丙烯酰胺凝胶在不同电压条件下电泳,2 g/L硝酸银染色,可以直接观察到大小不同的电泳条带,扩增的片段大多数在500 bp以下.研究表明,60g/L聚丙烯酰胺凝胶90 V电压电泳,电泳条带之间具有适当的距离,有利于差异条带分离和回收,是一种分离和检测DNA的有效方法.

Detection of the Products of Single Cow Embryo RT-PCR with Silver Staining PAGE

Silver staing PAGE has advantage to detect RT-PCR amplification products of early embryonic genes.In this study,the single embryo cultured in vitro was selected,RT-PCR amplification was carried out with anchor primer and random primer,and the products were detected with the different density polyacrylamide gel electrophoresis under the different electric voltage conditions.After 2 g/L silver nitrate dyeing,finally indicated gel electrophoresis belt can be observed directly,and majority products are below 500 bp,60 g/L polyacrylamide gel and 90 V voltage are good condition.The gel electrophoresis belts were separated in suitable distances,it was easy to recycle the difference gel electrophoresis belts.Gel electrophoresis is a kind of effective method for separatings and detecting DNA.