Figure 2. Primer design for PIGR ORF cloning by GDS strategy.

A. Exon primers for exon PCRs. All exons are represented by solid arrows and are flanked by optimized forward and reverse primers located in introns. The start codon resides in exon 2 and stop codon in exon 11. Exon 1 does not contain a coding sequence and was not amplified. For exon 11, only the region containing the coding sequence was amplified. B. Overlapping primers for overlapping PCR. C. Alignment of the overlapping primers with the cDNA sequence (only a portion is shown). Primers are boxed and their orientations are indicated by parallel arrows. Exon-exon junctions are marked by vertical lines.