Oligo(dT)23, Anchored primers are used to prime mRNA with a poly(A) tail for cDNA synthesis. The primers have 23 thymidine residues and one G, C, or A residue (the anchor) at the 3′ end. This anchor ensures that the oligo(dT) primer binds at the very beginning of the message and there is not a long region of the unusable sequence.
One approach to the analysis of gene expression is to measure the concentration of mRNA of a gene. There are several challenges to such analyses, such as the differences in half life between different transcripts, the temporal patterns of transcription and the lack of correlation between mRNA and protein.
The 3’/5’ integrity assay is a potential first step in the identification of RNA degradation. The assay is particularly useful when a large number of samples are to be analyzed or when the degradation is less than that detected by capillary systems but still sufficient to effect qPCR analyses.