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过量DMSO显著降低低模板浓度PCR扩增的特异性
引用本文:肖志壮,曲音波,汪天虹,高培基,刘梦海XIA Zhi-zhuang,QU Yin-bo,WANG Tian-hong,GAO Pei-ji,LIU Meng-hai.过量DMSO显著降低低模板浓度PCR扩增的特异性[J].遗传,2001,23(4):341-799.
作者姓名:肖志壮  曲音波  汪天虹  高培基  刘梦海XIA Zhi-zhuang  QU Yin-bo  WANG Tian-hong  GAO Pei-ji  LIU Meng-hai
作者单位:山东大学微生物技术国家重点实验室,济南 250100 State Key Laboratory of Microbial Technology,Shandong University,Jinan 250100,China
摘    要:DMSO通常经验性地用于提高PCR扩增的效率,但是过量的DMSO可以显著降低特异序列的扩增效率,尤其是导致非特异性扩增的现象却被忽视.在6%的DMSO存在时,开始出现非特异条带,同时特异扩增产物减少.本文首次报道DMSO对PCR产物特异性的影响并确定了克服上述现象产生的途径,即通过增加模板-引物的比例消除非特异条带.而通常提高复性温度只能部分减少非特异产物,不能避免非特异扩增.本文结果有助于提高常规PCR,尤其是分子遗传学分析中经常使用的随机扩增多态性DNA(randomamplifiedpolymorphicDNA,RAPD)检测的准确度。 Abstract:DMSO is the most often empirically used to increase efficiency of PCR.However,it is ignored that DMSO can also decrease the amplification of specific sequences and result in non- specific amphfications.Non-specific bands are detected at 6% DMSO,as specific amplification declines.This is the first report about the effects of DMSO on the specificity of PCR and establishment of the method for avoiding non-specific products,which can be overcome by increasing template-primer ratio rather than annealing temperature.Our data will be of much help for random amphfied polymorphic DNA (RAPD) assays for molecular genetic analyses.

关 键 词:D  M  SO  PCR  PCR  特异性和效率  Key  words  DMSO  

Excessive DMSO Dramatically Reducing the Specificity of PCR Amplification at Low Concentrations of Template
XIA Zhi-zhuang,QU Yin-bo,WANG Tian-hong,GAO Pei-ji,LIU Meng-hai.Excessive DMSO Dramatically Reducing the Specificity of PCR Amplification at Low Concentrations of Template[J].Hereditas,2001,23(4):341-799.
Authors:XIA Zhi-zhuang  QU Yin-bo  WANG Tian-hong  GAO Pei-ji  LIU Meng-hai
Abstract:
Keywords:
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