| 提高SCA2编码区CAG三核苷酸重复的PCR扩增效率 |
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| 引用本文： | 汤熙翔,夏家辉.提高SCA2编码区CAG三核苷酸重复的PCR扩增效率[J].生命科学研究,2000,4(3):217-221. |
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| 作者姓名： | 汤熙翔 夏家辉 |
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| 作者单位： | 湖南医科大学医学遗传学国家重点实验室，中国湖南长沙 410078 |
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| 基金项目： | 国家自然科学基金（39700080） |
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| 摘 要： | 以遗传性脊髓小脑共济失调Ⅱ型基因（ｓｐｉｎｏｃｅｒｅｂｅｌｌａｒ ａｔａｘｉａ ｔｙｐｅⅡｇｅｎｅＳＣＡ２）编码区内的ＣＡＧ三核苷酸重复为研究对象（Ｇ＋Ｃ含量为６９．２％），比较了热启动ＰＣＲ、碱基替代ＰＣＲ、添加增效剂（１％－１２．５％二甲亚砜、１％－２５％甘油、１％－１２．５％甲酰胺）与常规ＰＣＲ的扩增效率，发现热启动ＰＣＲ、碱基替代ＰＣＲ及添加增效剂（１％－１０％二甲亚砜、５％－２０％甘油、
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| 关 键 词： | 聚合酶链式反应 遗传性脊髓小脑共济失调Ⅱ型 |
| 文章编号： | 1007-7847（2000）03-0217-05 |
| 修稿时间： | 2000年8月8日 |
Efficacy Improvement of PCR Amplification of CAG Trinucleotide Repeats in the Coding Sequence of Spinocerebellar Ataxia Type II Gene |
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| TANG Xi-xiang,XIA Jia-hui.Efficacy Improvement of PCR Amplification of CAG Trinucleotide Repeats in the Coding Sequence of Spinocerebellar Ataxia Type II Gene[J].Life Science Research,2000,4(3):217-221. |
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| Authors: | TANG Xi-xiang XIA Jia-hui |
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| Abstract: | To improve the efficacy of PCR amplification of CAG trinucleotide repeats in the coding sequence of spinocerebellar ataxia type II gene(69.2％ G＋C）, hot-start PCR, base-replacement PCR, and the addition of enhancers(1％～12.5% DMSO , 1％～25% glycerol ,1％～12.5% formamide) were performed and compared with normal PCR . The results showed that hot-start PCR, base-replacement PCR and the addition of enhancers(1%～10% DMSO , 5%～20% glycerol , 1%～10% formamide) improved the amplification efficacy of the GC rich region. Gene diagnosis in 70 SCA pedgrees and 60 spontaneous SCA patients were also conducted. |
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| Keywords: | polymerase chain reaction spinocerebellar ataxia type II amplification efficacy |
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