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. Author manuscript; available in PMC: 2009 Jun 22.
Published in final edited form as: Mol Biochem Parasitol. 2007 Nov 22;158(1):32–45. doi: 10.1016/j.molbiopara.2007.11.009

Figure 1. Schematic of TAP/EpiTAP PCR Reaction.

Figure 1

TAP and Epi-TAP PCR products were generated as instructed by the manufacturer (Gene Therapy Systems) using their TAP and Epi-TAP Express™ kits but modified for Plasmodium using a two step PCR reaction. In the first step, primers were designed incorporating a universal sequence for either a TAP or Epi-TAP amplification with the Epi-TAP primer including an HA epitope tag. The 5′ primers begin with one of these universal sequences and continue with a start codon ATG which is followed by 15–20 bp of the 5′ sequence of the Plasmodium spp. gene of interest. The 3′ primers for both TAP and EpiTAP begin with the same universal sequence and continue with a stop codon TCA followed by 15–20 bp’s of the anti-sense terminal sequence of the Plasmodium sp. gene. The second TAP and EpiTAP reactions used a 1:20 dilution of the first step product, and proprietary CMV promoter and SV40 terminator fragments as templates. The promoter and terminator fragments overlap with the first step PCR product at, respectively, 5′ and 3′ end of the universal region and, as the result of the second step PCR, the promoter, the gene of interest and the terminator are linked in the correct order to form a transcriptionally active PCR (TAP) fragment.