[Optimization of hybridization efficiency in cDNA chip technology].
OBJECTIVE
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.


METHODS
Factors are changed one by one. Optimal values are selected by comparing those of different groups.


CONCLUSIONS
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.
Ma
Shu-hua
SH
Academy of Military Medical Sciences, Institute of Radiation Medicine, Beijing 100850, China.
Wang
Sheng-qi
SQ
D001923:Brain Chemistry; D018076:DNA, Complementary / Q000737:chemistry*; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification
D018076:DNA, Complementary; D012313:RNA