Effects of additives on efficiency and specificity of ligase detection reaction.
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.
Xiao
Zhen-Xian
ZX
State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, Shanghai 200237, PR China.
Cao
Hui-Min
HM
Luan
Xiao-Hui
XH
Zhao
Jian-Long
JL
Wei
Dong-Zhi
DZ
Xiao
Jun-Hua
JH
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
D015342:DNA Probes; D005559:Formamides; D007202:Indicators and Reagents; D011136:Polysorbates; D011092:Polyethylene Glycols; C031066:formamide; D005990:Glycerol; D004121:Dimethyl Sulfoxide