How can I test if blocking of cDNA synthesis is specific for globin mRNA transcripts?
The specificity of the method can be tested by using the GeneChip Globin Reduction RNA Controls kit which contains two control samples, Jurkat RNA and Jurkat RNA + Globin RNA. These controls mimic blood RNA samples not affected by excess globin message (for example, an erythrocyte lysis fractionation method) and affected by excess globin (for example, a PAXgene blood RNA sample) respectively. Besides providing additional possibilities for validation of the globin reduction procedure, target preparation starting from Jurkat RNA (which is free of globin transcripts) can be performed with and without integrated globin reduction. Thus, specificity can be confirmed by comparing the expression profiles obtained from each sample. If non-globin transcripts are unaffected by the globin reduction procedure, no significant differences in the expression profiles derived is expected. For more information on the GeneChip Globin Reduction RNA Controls kit, please refer to the GeneChip Globin-Reduction Kit handbook.