IRE-bubble PCR: a rapid method for efficient and representative amplification of human genomic DNA sequences from complex sources.
IRE-bubble PCR: a rapid method for efficient and representative amplification of human genomic DNA sequences from complex sources.
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DOI：
10.1006/geno.1994.1100
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发表时间：
1994-02
期刊：
影响因子：
4.4
通讯作者：
D. Munroe;M. Haas;E. Bric;Tania Whitton;H. Aburatani;K. Hunter;D. Ward;D. Housman
中科院分区：
文献类型：
--
作者：
D. Munroe;M. Haas;E. Bric;Tania Whitton;H. Aburatani;K. Hunter;D. Ward;D. Housman
分析任何基因组DNA片段的一个重要问题是产生一组代表该区域的独特的短单拷贝序列。在这份报告中，我们描述了一种新的技术，IRE-bubble PCR，它被设计成扩增体细胞杂交，YACs，cosmetic，和λ噬菌体的人类DNA含量，并导致更大的复杂性和代表性比标准的IRE间PCR。在这里，我们证明了IRE气泡PCR是物种特异性的，它导致产生的产物比标准IRE间PCR产生的产物复杂和具有代表性至少10倍。此外，我们还讨论了影响IRE气泡PCR产物表达的因素，并展示了它们如何用于进一步增加该反应的复杂性。最后，我们已经说明了如何通过IRE气泡PCR产生的产物的复杂性和分布可以被利用和应用于FISH映射和“染色体绘画”以及产生针对特定染色体或亚染色体区域的STS。
A significant issue in the analysis of any genomic DNA segment is the generation of a unique set of short single-copy sequences that are representative of that region. In this report we describe a novel technique, IRE-bubble PCR, which was designed to amplify the human DNA content of somatic cell hybrids, YACs, cosmids, and lambda phage and result in greater complexity and representation than standard inter-IRE PCR. Here we demonstrate that IRE-bubble PCR is species specific and that it results in the generation of a product that is at least 10-fold more complex and representative than that produced by standard inter-IRE PCR. In addition, we have addressed the factors that contribute to the representation of the IRE-bubble PCR product and show how they may be used to further increase the complexity of this reaction. Finally, we have illustrated how the complexity and distribution of products generated by IRE-bubble PCR can be exploited and applied to FISH mapping and "chromosome painting" as well as to the generation of STSs targeted to specific chromosomal or subchromosomal regions.