Abstract:Gene chip was prepared with the oligonucleotide probes of 6 virulence genes of Vibrio anguillarum dot-applied on a piece of positively charged nylon membrane using the microcaster manual arrayer, followed by cross-linkage under the ultraviolet light. The detection of V. anguillarum virulence genes was performed by hybridizing with the DIG-labeled PCR products on the membrane chip. Here we report the optimization of the concentration of the probe, the time course of hybridization, and the process of washing steps. After optimization, the hybridization time was reduced to 30min, the concentration of the probe as 50μmol/L, and the process of washing was shortened to 3 steps of only 45 min.
