Analysis of cDNA subtraction. (A) Electrophoresis of PCR products of subtracted small and large cDNA. After the second round of suppressive PCR, 5 μL product was run on a 2.0% agarose gel. To obtain clear resolution of fine bands, the gel was run at 80 V. DNA size markers (bp) are indicated for the 1-kb DNA ladder in lane M. (B) Comparison of subtracted and nonsubtracted cDNA for small and large LTC-DCs. Subtraction efficiency was judged by comparing the prevalence of the housekeeping gene GAPDH before and after subtraction. After 18, 23, 28, or 33 cycles of PCR using GAPDH 3′ and 5′ primers, 5 μL PCR product was run on a 2.0% agarose gel.