Figure 3
Qualitative RT-PCR analysis of HEK293T cells transfected with pcDNA3.1-DGAT1 (WT or M435L) constructs. (A) Illustration of plasmid constructs used for qualitative RT-PCR. pcDNA3.1-DGAT1 (WT) and pcDNA3.1-DGAT1 (M435L) are carrying alternatively the p.M435 or the p.L435 alleles, respectively. Blue boxes and lines represent bovine DGAT1 exons and introns, respectively. Black line represents pcDNA3.1 (+) backbone. The P5-P2 primer pair and its target region is indicated in red. (B) Gel electrophoresis of P5-P2 RT-PCR products obtained from HEK293T cells transfected with pcDNA3.1-DGAT1 (WT or M435L). The structure of amplicons 1 to 3 is depicted on the right. NT, non-transfected ; RT (-), RT-PCR performed without Superscript III. (C) Validation of amplicon structure from (B) by Sanger sequencing. Only exon-intron and exon-exon boundaries are shown.
