Effects of additives on efficiency and specificity of ligase detection reaction

• Published in: Molecular biotechnology Vol. 35; no. 2; pp. 129 - 133
• Main Authors: XIAO, Zhen-Xian, CAO, Hui-Min, LUAN, Xiao-Hui, ZHAO, Jian-Long, WEI, Dong-Zhi, XIAO, Jun-Hua
• Format: Journal Article
• Language: English
• Published: Totowa, NJ Humana Press 01.02.2007; Springer Nature B.V
• Subjects: Additives; Base Sequence; Biological and medical sciences; Biotechnology; Dimethyl Sulfoxide; DNA Probes - genetics; Enzymes; Formamides; Fundamental and applied biological sciences. Psychology; Glycerol; Humans; Indicators and Reagents; Ligase Chain Reaction - methods; Ligase Chain Reaction - statistics & numerical data; Molecular biology; Molecular Sequence Data; Polyethylene Glycols; Polysorbates; Sensitivity and Specificity; Studies; Specificity; additives; Enzyme; Efficiency; Ligases; LDR; Detection
• ISSN: 1073-6085; 1559-0305
• DOI: 10.1007/BF02686107

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.