FIG. 3.

Requirement for proximal chaperone. Two micrograms of G. chapellei total RNA was hybridized overnight in 4× SSPE–2.5× Denhardt's solution–30% formamide buffer at room temperature to the microarray targeting positions 214 and 420 (Fig. 1). Detector probes located at different positions with respect to the capture oligonucleotide were hybridized simultaneously with the target RNA and microarray slide. Positive hybridization was detected with the Molecular Probes chemiluminescent substrate ELF-97and a Bio-Rad Fluor-S imager. Relative light units (OD ± standard error) were measured with Phoretix software. Results indicate that the chaperone-detector probe should bind immediately adjacent to the capture oligonucleotide, as shown in Fig. 2D.