Uniform amplification of multiple DNAs by emulsion PCR
Uniform amplification of multiple DNAs by emulsion PCR
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DOI：
10.1016/j.bbrc.2006.11.037
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发表时间：
2007-01-12
影响因子：
3.1
通讯作者：
Suzuki, Yosuke
中科院分区：
文献类型：
--
作者：
Hori, Machiko;Fukano, Hajime;Suzuki, Yosuke
当在一个PCR管中通过PCR扩增几个DNA时，已知会发生偏倚扩增，因为每个DNA的扩增效率不同。因此，我们在油包水乳液（W/O乳液）中进行PCR，以检查该程序是否允许几种DNA的均匀扩增。在由两个克隆组成的模型文库的扩增中，PCR混合物的乳化成功地将其扩增效率的差异降低到没有乳化时获得的值的约七分之一。此外，我们进行了重复PCR扩增由10个短发夹RNA（shRNA）表达载体组成的模型文库，作为使用shRNA表达文库进行基因发现的模型实验。因此，PCR混合物的乳化成功地降低了PCR偏倚。我们的研究结果表明，乳液PCR能够均匀扩增的shRNA，核酶，cDNA和其他的文库，也是有用的基因发现使用这些库。(c)2006年爱思唯尔公司All rights reserved.
When several DNAs are amplified by PCR in one PCR tube, biased amplification is known to occur because amplification efficiency differs from one DNA to another. Therefore, we conducted PCR in the water in oil-emulsion (W/O emulsion) to examine whether the procedure allows the uniform amplification of several DNAs. In the amplification of a model library consisting of two clones, the emulsification of the PCR mixture successfully reduced the difference in its amplification efficiency to approximately one-seventh the value obtained without emulsification. Furthermore, we conducted repeated PCR to amplify a model library consisting of ten short hairpin RNA (shRNA) expression vectors as a model experiment for gene discovery using an shRNA expression library. Consequently, the emulsification of the PCR mixture successfully reduced PCR bias. Our results indicate that emulsion PCR is capable of uniformly amplifying libraries of shRNA, ribozyme, cDNA, and others, and is useful also for gene discovery using these libraries. (c) 2006 Elsevier Inc. All rights reserved.