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. 2003 Feb 6;72(3):749–758. doi: 10.1086/368263

Figure 1.

Figure 1

IKBKAP amplicon sizes and their corresponding PCR products. A schematic representation of the primer sets and Taqman probes used to amplify either the WT or MU IKBKAP transcripts. PCR primers 19F–23R amplify both WT and MU bands. The real-time QPCR primers designated “WT-F/R” (forward/reverse) or “MU-F/R,” with their matching dual-labeled Taqman probes, amplify WT and MU transcripts, respectively. The position of WT-R primer in exon 20 was designed to specifically amplify only the WT message, whereas the MU-F primer located at the junction of exons 19 and 21 will amplify only the MU message. The dual-labeled Taqman probes have a melting temperature (Tm) of 7°C–10°C above the primer Tm for each amplicon. Primer and probe sequences were as follows: 19F: CCT GAG CAG CAA TCA TGT G; 23R: TAC ATG GTC TTC GTG ACA TC; WT-F: GCA GCA ATC ATG TGT CCC A; WT-R: ACC AGG GCT CGA TGA TGA A; WT Taqman probe: FAM-GTT CAC GGA TTG TCA CTG TTG TGC C-BHQ; MU-F: CAC AAA GCT TGT ATT ACA GAC T; MU-R: GAA GGT TTC CAC ATT TCC AAG; and MU Taqman probe: FAM-CTC AAT CTG ATT TAT GAT CAT AAC CCT AAG GTG-BHQ. The housekeeping gene 18S rRNA was used for normalization: 18S-F: GGC CCT GTA ATT GGA ATG AG; 18S-R: GCT ATT GGA GCT GGA ATT AC; and 18S Taqman probe: FAM-TGC TGG CAC CAG ACT TGC CCT C-BHQ.