| PAGE凝胶电泳银染方法的改进 |
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| 引用本文： | 张金成,梁永文,范丽娜,廖和荣.PAGE凝胶电泳银染方法的改进[J].中国畜牧兽医,2009,36(7):99-101. |
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| 作者姓名： | 张金成 梁永文 范丽娜 廖和荣 |
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| 作者单位： | (石河子大学动物科技学院， 石河子 832003) |
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| 基金项目： | 国家自然科学基金，石河子大学大学生研究训练计划项目 |
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| 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pBR322 DNA/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25 ng，但在染色所需时间和可操作性方面，改进的染色方法优于Sanguinetti等所建立的方法。改进的银染方法染色过程较快，仅需25 min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。
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| 关 键 词： | 微卫星 聚丙烯酰胺凝胶电泳 银染 |
Advanced Technique for Silver Staining Methods on Polyacrylamide Gels Electrophoresis |
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| ZHANG Jin-cheng,LIANG Yong-wen,FAN Li-na,LIAO He-rong.Advanced Technique for Silver Staining Methods on Polyacrylamide Gels Electrophoresis[J].China Animal Husbandry & Veterinary Medicine,2009,36(7):99-101. |
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| Authors: | ZHANG Jin-cheng LIANG Yong-wen FAN Li-na LIAO He-rong |
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| Institution: | (College of Animal Science and Technology， Shihezi University， Shihezi 832003， China) |
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| Abstract: | 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 MspⅠDNA molecular weight. 2 silver staining methods were analyzed and evaluated. The sensitivity of all the band of pBR322 MspⅠ 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． |
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| Keywords: | microsatellite polyacrylamide gel electrophoresis silver staining |
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