15 mer poly dt oligonucleotide

Organs were removed from immunized mice and lysed using a homogenizer. Total RNA was extracted from lysates in the presence of RNase inhibitors using TRIzol reagent (Molecular Research Center, Cincinnati, OH, USA). Isolated RNA was dissolved in diethyl pyrocarbonatetreated water (Sigma-Aldrich, St. Louis, MO, USA) and used to generate cDNA using a 15mer poly dT oligonucleotide (Invitrogen, Carlsbad, CA, USA) and Superscript reverse transcriptase (Gibco) with incubation at 37°C for 1 h according to the manufacturer's protocol.
Expression of the hNIS gene was detected in dLNs, non-dLNs, and spleen by RT-PCR using the primers: 5'-GGCTCCTCGGTGACTCTAGGATGC-3' (forward) and 5'-CATGAATTCTGGGCTCAATTTTCTTGTCC-3' (reverse). To confirm DNA integrity, the mouse β-actin gene (codons 135-223) was amplified with the primers 5'-GGCTCCTCGGTGACTCTAGGATGC-3' (forward) and 5'-CATGAATTCTGGGCTCAATTTTCTTGTCC-3' (reverse) under the following conditions: 34 cycles of 94°C at 60 s, 55°C at 60 s, and 72°C at 60 s.