| 2种方法银染的AFLP片段再扩增效果比较及改进 |
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| 引用本文： | 刘耘,熊家军,杨利国.2种方法银染的AFLP片段再扩增效果比较及改进[J].华中农业大学学报,2008,27(2):182-185. |
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| 作者姓名： | 刘耘 熊家军 杨利国 |
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| 作者单位： | 华中农业大学动物遗传育种与繁殖教育部重点实验室,武汉,430070 |
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| 基金项目： | 国家高技术研究发展计划(863计划) |
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| 摘 要： | Bassam银染法和Sanguinetti银染法是AFLP标记最常用的检测方法.本研究比较这2种方法银染的AFLP条带再扩增效果的差异,发现Bassam法银染的AFLP片段再扩增效率达到100%,而用同样的PCR条件,sanguinetti法银染的AFLP片段不能被扩增,限制了Sanguinetti法在AFLP标记分析中的应用.为提高Sanguinetti法银染的AFLP片段再扩增效率,对Sanguinetti法银染的AFLP片段回收和再扩增条件进行了改进.通过增加Sanguinetti法显色后的胶板漂洗次数、延长漂洗时间、改变AFLP条带浸泡溶液、增加浸泡液体积和增加PCR循环次数,成功地将Sanguinetti法银染的AFLP条带的再扩增效率提高到100%,从而解决了San-guinetti银染法应用的限制因素.
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| 关 键 词： | AFLP 银染 Bassam法 Sanguinetti法 再扩增 |
Comparison of the Re-amplification Effects of AFLP Bands Stained with Two Silver Staining Methods and Improvement |
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| LIU Yun XIONG Jia-jun YANG Li-guo.Comparison of the Re-amplification Effects of AFLP Bands Stained with Two Silver Staining Methods and Improvement[J].Journal of Huazhong Agricultural University,2008,27(2):182-185. |
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| Authors: | LIU Yun XIONG Jia-jun YANG Li-guo |
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| Institution: | Key Laboratory of Agricultural Animal Genetics;Breeding and Reproduction of Ministry of Education;Huazhong Agricultural University;Wuhan 430070;China |
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| Abstract: | The Bassam and Sanguinetti methods are the most frequently used silver staining techniques for the detection of AFLP marker at present.In this study,we compared the efficiency of the re-amplification of AFLP bands recovered from polyacrylamide gels stained with the Bassam and Sanguinetti methods.The re-amplification success rate of AFLP bands recovered from gels stained with the Bassam method reaches 100%,while that of the Sanguinetti gels is very low.Low level of re-amplification success rate restricts the... |
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| Keywords: | AFLP silver staining Bassam method Sanguinetti method re-amplification |
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