Figure 4. Detection of amplified products corresponding to KV1.1–1.6 mRNA in rat brain and cerebral arteries.
A, RT-PCR screening detected mRNA encoding KV1.1–1.6 α-subunits in lanes loaded with amplified cDNA originating from rat brain (Br, left lanes) or rat middle cerebral arteries (C, right lanes). Detection of each product was verified in cDNA reverse transcribed from 7–12 different RNA isolations. Each isolation was prepared from tissues of two rats. The brain was used as a positive control. Expected product sizes (bp) were: 284 (1.1), 374 (1.2), 581 (1.3), 643 (1.4), 1111 (1.5), 578 (1.6). B, screening for genomic contamination of cDNA using primers for smooth muscle-specific α-actin that included three introns in the amplified region. Amplification reactions including cDNA (70 nmol) resulted in a 637 bp product consistent with the mRNA template, whereas reactions including DNA (70 nmol) resulted in the predicted 1506 bp product amplified from the intron-containing region. Control reactions included amplifications without cDNA or without DNA. Results were verified in at least three different preparations. Each preparation was obtained from tissues of two rats.