Optimization of gene expression analysis using immobilized capture probes of different lengths and densities

摘要

Disclosed is a method of obtaining similar signal intensities from hybridization of different sets of the oligonucleotide probes to a set of cognate targets, in a hybridization assay that comprises sets of probes of different lengths on different solid phase carriers and sets of targets of different lengths, wherein annealing of a set of probes with cognate targets generates a signal intensity, wherein a cumulative signal intensity from said annealing is determined, the method comprising: (i) selecting probe lengths and densities for said sets of probes, wherein probes include different subsequences such that at least one subsequence is complementary to a subsequence in a cognate target; wherein probes for longer cognate targets are longer in length than probes for shorter cognate targets and wherein the density for longer probes is lower than the density for shorter probes, (ii) producing said probes and affixing said probes to different solid phase carriers at a selected density, (iii) annealing said target to said probes, wherein signal intensities of probes and targets of different lengths are about the same.