一种新的用于提高lea3基因扩增特异性和产量的增强剂(英文)  

Effect of New Specificity and Yield Enhancer on Amplification of lea 3 Gene

作　　者：于丽霞[1] 武晓璐[1] 汤晓倩[1] 鄢波[1] 

机构地区：[1]西南林业大学园林学院,云南昆明650224

出　　处：《云南植物研究》2010年第6期535-538,共4页Acta Botanica Yunnanica

基　　金：The Applied Foundation Research of Yunnan Province in China (Project number 2009ZC079 M);Yunnan Province and Minister Key Subject, University Key Lab and Share Platform

摘　　要：在对目的DNA序列尤其是高GC含量的片段进行PCR扩增的时候,经常需要对一些试验条件进行优化。在一定条件下,二甲基亚砜、甲酰胺、甘油、NP-40和Tween20等可以在某种程度上提高PCR的特异性和效率。我们在对禾本科lea3基因进行分离克隆时发现了一种新的可以提高PCR产量和特异性的物质——极高热稳定单链结合蛋白(ETSSB),研究发现在每50μlPCR反应体系中加入200ng的ETSSB,可以有效地抑制DNA片段的非特异性条带的产生,并可以提高目的片段的产量。Amplification of a target DNA by the polymerase chain reaction （PCR） often requires experimental optimization factors especially when the target sequence contains high content of GC. Dimethyl sulfoxide,formamide,glycerol,NP-40 and Tween20 are known to increase,the specificity and efficiency of PCR under certain conditions. In the amplification of lea3 gene in Gramineae,we found a new Extreme Thermostable Single-Stranded DNA Binding Protein （ET SSB） and improved yield of PCR. Addition of 200 ng of ET SSB per 50 μl reation decreased the formation of non-specific DNA fragments and increased the yield of specific PCR products.

关 键 词：高GC含量 聚合酶链式反应 极高热稳定单链结合蛋白 

分 类 号：Q943.2[生物学—植物学]