同源引物的非等量PCR
Henan Science(2018)
Abstract
为确定引物量对同源引物扩增DNA的影响.将正向引物GFO与同源度为45%、90%、100%、54%、9%的反向引物CR1、CR2、CR3、CR4、CR5分别用于扩增5.9 kb pET20b-C2-G10-C2模式DNA.等量引物(500 nmol/L)时, CR3和CR4均无法扩增目的条带.而非等量PCR时,将反向引物量降低10倍(50 nmol/L),CR3和CR4均扩增出目的条带,即正向引物量降低10倍时,五对引物均扩增出目的条带.CR3降低100倍、CR4降低10倍扩增DNA最多,经15~20次循环五对引物扩增DNA最多.实验结果表明,非等量PCR通过降低单侧引物量减少引物二聚体形成,从而扩增双链DNA.
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Key words
unequal quantity,complementary primer,PCR,DNA
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