3'Seq was designed to quantitatively measure the abundance of 3' untranslated regions (UTR) isoforms in a wide-array of human tissue samples types. The first cDNA strand is generated by reverse transcribing total RNA using an oligo (dT) primer containing a VN-anchor (where V is dA, dC, or dG), P5 adapter sequence, a uridine, and biotin that is bound to a streptavidin-coated magnetic bead. The second cDNA strand is synthesized using DNA polymerase I. To initiate nick translation with DNA polymerase I, RNase HII was used to introduce a nick at the uridine. This creates another nick that is 50-75 bases away from the 3' end. A blunt end is created at the position of the new nick and double-stranded P7 adapters are ligated at this position. The double-stranded cDNA is PCR amplified, purified, and ready to be sequenced.