摘要
为改良DNA聚丙烯凝胶电泳(PAGE)银染方法,采用PCR方法扩增鸡基因组微卫星DNA序列后,经PAGE分离扩增产物和pBR322DNA/BsuRI标准分子质量,利用2种银染法对微卫星位点进行检测,分析其灵敏度和可行性。结果表明,2种方法都能染出Marker的所有条带,灵敏度均为25ng,但在染色所需时间和可操作性方面,改进的染色方法优于San-guinetti等所建立的方法。改进的银染方法染色过程较快,仅需25min左右,且染色液和显影液可回收利用。该方法能显著提高检测分辨率,在实际应用中提高了染色效率。
To study and modify the optimal methods of silver staining of DNA polyacrylamide gels electrophoresis, polymerase chain reaction （PCR）-polyacrylamide gel electrophoresis silver nitrate （AgNO3） staining were used to detect DNA fragments and pBR322 Msp I DNA molecular weight. 2 silver staining methods were analyzed and evaluated. The sensitivity of all the band of pBR322 Msp I Marker developed was coequally 25 ng. However, comparison of time and step,advanced technique was more convenient. The advanced staining cost only about 25 minutes, and was characterized as simple steps,high sensitivity, little pollution, which was recycled and valuable for practical apply.
出处
《中国畜牧兽医》
CAS
北大核心
2009年第7期99-101,共3页
China Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金项目(30560101)
石河子大学大学生研究训练计划项目(SRP0714243)
关键词
微卫星
聚丙烯酰胺凝胶电泳
银染
microsatellite
polyacrylamide gel electrophoresis
silver staining
