2.3. Alternative Splicing of HLA-DPA1 (Experiment 3)
By exon array, a variable expression pattern was observed across exons 2, 3, and 4 in HLA-DPA1 for multiple subjects. This gene was therefore chosen for analysis of alternative splicing by direct sequencing of gel-purified cDNA amplicons to evaluate potential variants.
Representative cDNA samples from subjects in Experiment 2 were PCR amplified and run on agarose gels for separation of bands. cDNA amplification was conducted from exons 2 through 4, to screen for alternative splicing. Gel bands smaller than full length were visualized on gels, and sent for Sanger sequencing. The subjects screened are shown in Supplementary Table 1.