Long oligonucleotide arrays on nylon for large-scale gene expression analysis.
To date, most studies of multigenic expression patterns by long DNA array have used DNA fragments as probes. These probes are usually obtained as PCR products, and this represents a time-consuming and error-prone approach, requiring strict quality control. The present study examines the use of 40- and 70-mer synthetic oligonucleotides as probes for DNA array analysis with radioactive labeled targets. Design, spotting onto nylon filters, and hybridization conditions were determined and optimized. In this approach, the sensitivity and the specificity of the hybridization appear comparable to the conventional long DNA probes assay, permitting the analysis of small samples of approximately 1 microg total RNA. The long oligonucleotide array thus provides a very convenient method for the analysis of gene expression patterns in biological specimens and in clinical research.
El Atifi
M
M
CHRU, Equipe Transcriptome, INSERM U318, Laboratoire de Neurosciences Précliniques, Grenoble, France. melatifi@chu-grenoble.fr
Dupré
I
I
Rostaing
B
B
Chambaz
E M
EM
Benabid
A L
AL
Berger
F
F
D017360:Arabidopsis / Q000235:genetics; D015342:DNA Probes; D005240:Feasibility Studies; D020869:Gene Expression Profiling / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D007553:Isotope Labeling / Q000379:methods; D008422:Materials Testing; D008567:Membranes, Artificial; D009757:Nylons; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods*; D010759:Phosphorus Isotopes; D011786:Quality Control; D012313:RNA / Q000032:analysis / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D015342:DNA Probes; D008567:Membranes, Artificial; D009757:Nylons; D010759:Phosphorus Isotopes; D012313:RNA