title: Effects of additives on efficiency and specificity of ligase detection reaction.
abstract: Ligase detection reaction (LDR) is adaptable to a wide variety of applications ranging from scientific research to clinical diagnosis, especially in the field of nucleotide polymorphism discrimination and analysis. Efficiency and specificity of LDR are the most two important characteristics that influence its application. To improve the specificity or efficiency of ligase, optimization of the design of LDR probes and the reaction of LDR were investigated previously by most researchers. But the effects of additives on LDR have not been reported. In this study, the effects of additives (DMSO, Tween-20, glycerol, formamide, and PEG- 6000) on LDR efficiency and specificity were investigated. The results showed that all of these compounds, except for Tween-20, could improve the specificity of LDR. PEG-6000 was proved to be the best additive among the five tested with an optimal concentration of 5% at which the highest yield was obtained with a relatively improved specificity.
authors: State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, Shanghai 200237, PR China.
authors: Zhen-Xian
authors: ZX
authors: Xiao
authors: Hui-Min
authors: HM
authors: Cao
authors: Xiao-Hui
authors: XH
authors: Luan
authors: Jian-Long
authors: JL
authors: Zhao
authors: Dong-Zhi
authors: DZ
authors: Wei
authors: Jun-Hua
authors: JH
authors: Xiao
mesh_terms: D001483:Base Sequence; D001709:Biotechnology; D015342:DNA Probes / Q000235:genetics; D004121:Dimethyl Sulfoxide; D005559:Formamides; D005990:Glycerol; D006801:Humans; D007202:Indicators and Reagents; D022521:Ligase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D008969:Molecular Sequence Data; D011092:Polyethylene Glycols; D011136:Polysorbates; D012680:Sensitivity and Specificity
chemical_list: D015342:DNA Probes; D005559:Formamides; D007202:Indicators and Reagents; D011136:Polysorbates; D011092:Polyethylene Glycols; C031066:formamide; D005990:Glycerol; D004121:Dimethyl Sulfoxide