ROBUST AMPLIFICATION AND ETHIDIUM-VISIBLE DETECTION OF THE FRAGILE-X-SYNDROME CGG REPEAT USING PFU POLYMERASE
ROBUST AMPLIFICATION AND ETHIDIUM-VISIBLE DETECTION OF THE FRAGILE-X-SYNDROME CGG REPEAT USING PFU POLYMERASE
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DOI：
10.1002/ajmg.1320510447
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发表时间：
1994-07-15
期刊：
影响因子：
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通讯作者：
HUGHES, MR
中科院分区：
文献类型：
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作者：
CHONG, SS;EICHLER, EE;HUGHES, MR
我们报告了溴化乙锭检测FMR 1正常和前突变大小的CGG三联体重复序列的方法。一组PCR条件已被优化的极端嗜热菌，Pyrococcus furiosus的聚合酶。使用该方案，可以通过琼脂糖凝胶的溴化乙锭染色检测范围为5至50个重复的正常大小的等位基因，以及范围为>50至约200个重复的大多数突变前大小的等位基因。由于该方案既不需要放射自显影，也不需要聚丙烯酰胺凝胶电泳，它有望提供一种快速的手段，排除脆性X综合征的男性精神发育迟滞。(C)1994 Wiley-Liss，Inc.
We report on a method for ethidium bromide detection of FMR1 normal and premutation-size CGG triplet repeats. A set of PCR conditions has been optimized for the polymerase of the hyperthermophilic bacterium, Pyrococcus furiosus. Using this protocol, normal-size alleles ranging from 5 to 50 repeats, as well as most of premutation-size alleles, varying from >50 to approximately 200 repeats, could be detected by ethidium bromide staining of agarose gels. Since the protocol requires neither autoradiography nor polyacrylamide gel electrophoresis, it promises to provide a rapid means for the exclusion of fragile X syndrome in males with mental retardation. (C) 1994 Wiley-Liss, Inc.