Optimization of hybridization efficiency in cDNA chip technology / 药学学报

ABSTRACT

<p><b>AIM</b>There are many factors affecting hybridization in cDNA chip DNA concentration immobilized on glass surface, spotting solution dissolving DNA, the concentration and purity of fluorescence-labeled probe, hybridization solution, hybridization temperature, hybridization time, Cy3- and Cy5-labeled probes, etc. In order to improve hybridization efficiency, tests were designed to optimize these factors.</p><p><b>METHODS</b>Factors are changed one by one. Optimal values are selected by comparing those of different groups.</p><p><b>RESULTS AND CONCLUSION</b>The efficient hybridization condition is as follows 0.50 microgram.microL-1 DNA fragments resolved in 1 x Micro Spotting Solution (ArrayIt) hybridize with purified fluorescent probe in 0.5% SDS/10 x SSC at 42 degrees C.</p>