Biphasic amplification of very dilute DNA samples via 'booster' PCR.
Biphasic amplification of very dilute DNA samples via 'booster' PCR.
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DOI：
10.1093/nar/17.13.5407
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发表时间：
1989-07
影响因子：
14.9
通讯作者：
G. Ruaño;W. Fenton;K. Kidd
中科院分区：
文献类型：
--
作者：
G. Ruaño;W. Fenton;K. Kidd
少于1000个拷贝的模板的聚合酶链式反应扩增会因为合成所谓的引物二聚体和其他假冒产物而受到阻碍。这些人工产物消耗了引物的库存，并占用了酶，从而降低了靶标的产量。为了克服这个问题，我们设计了Booster PCR，一种双相扩增策略。在第一阶段中，将引物稀释以获得初始107倍于模板的过量。在第二阶段的开始，将引物浓度提高到0.1M，这一过程不同于标准的PCR方法，后者从一开始就使用0.1-1.0#M的引物。其中扩增是从非常人类的基因组开始的，与PCR3非靶标扩增产物人工产物、分子拷贝(PG)、引物二聚体形成对比。副本
PCR amplification of fewer than 1000 copies of template is hampered by synthesis of so-called primer dimers and other spurious products. These artifacts consume the stock of primers and occupy enzyme, with consequent reduction in yield of target. To overcome this problem, we have devised Booster PCR, a biphasic amplification strategy. During Stage I, primers are diluted to obtain an initial 107-fold molar excess of primer to template. At the beginning of Stage II, primer concentration is brought up to 0.1 ;,M. This procedure differs from standard PCR methods which use 0.1-1.0 #M primers from the outset. which amplification started from very human genomic In contrast to PCR 3 non-target amplified products artifacts artifacts molecular copies pg), primer dimers. copies