Laser capture microdissection-generated target sample for high-density oligonucleotide array hybridization

Current advances in biomolecular technology allow precise genetic fingerpri nting of specific cells responsible for the pathogenesis of human diseases. This study demonstrates the feasibility of generating target samples from laser capture microdissection (LCM) tissues suitable for hybridization of h igh-density oligonucleotide arrays for gene expression profiling. RNA was s uccessfully isolated by LCM from three paired specimens of oval cancer and linearly amplified using T7 RNA polymerase. Evaluation of the cDNA revealed that five of five cellular maintenance transcripts are detected Biotinylat ed cRNA was generated and hybridized to the human Test I GeneChip(R) probe arrays, which demonstrated that the RNA is of sufficient quality and integr ity to warrant further analysis Subsequent hybridization of the samples to the HuGenFL GeneChip probe arrays revealed that 26.5%-33.0% of the approxim ately 7000 represented genes are expressed in each of the six samples. Thes e results demonstrate that LCM-generated tissues can generate sufficient qu ality cRNA for high-density oligonucleotide microarray analysis, an importa nt step in determining comprehensive gene expression profiling using this h igh-throughput technology.