Does the globin reduction procedure affect the expression analysis of transcripts different from globin?
The PNA oligomers are highly specific for globin mRNA transcripts and do not hybridize stably to non-globin transcripts under the recommended conditions. Therefore, following hybridization of the PNAs and Oligo d(T) Promoter Primer, amplification and labeling of non-globin transcripts should be unaffected. Since the reduction of excess labeled globin transcript in the final cRNA product significantly reduces the globin cross-hybridization effects, GeneChip analysis will be more sensitive. As detection of transcripts expressed at lower levels will no longer be masked by excess globin cross-hybridization, and the relative expression levels on a gene-by-gene basis will be higher.