DNA hybridization in reverse micelles and its application to mutation detection.
DNA hybridization was investigated in AOT (sodium di-2-ethylhexyl sulfosuccinate)/isooctane reverse micelles. The single-stranded DNA molecules were encapsulated in the nanoscale water pools formed in the reverse micelles, reducing the hybridization rate. The DNA hybridization can be monitored by simply measuring the UV absorbance of the reverse micellar solution at 260 nm. We found that the DNA hybridization occurred only at the restricted water content (Wo = [H2O]/[AOT] = 20) and below room temperature. We applied this DNA hybridization behavior in reverse micelles to mutation detection in a model gene p53 and successfully detected the single nucleotide mutations in 20-mer. 30-mer and 50-mer nucleotides without a DNA labeling.
Park
Lian-Chun
LC
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, 6-10-1 Hakozaki, Fukuoka 812-8581, Japan.
Maruyama
Tatsuo
T
Goto
Masahiro
M
D004252:DNA Mutational Analysis; D016158:Genes, p53; D008823:Micelles; D009693:Nucleic Acid Hybridization / Q000379:methods*; D013056:Spectrophotometry, Ultraviolet
D008823:Micelles