Fig. 2.

PCR products separated in a 2% agarose gel. Rows a-d depicts gradient PCR of exon 18, 19, 20 and 21 respectively. M denotes 100kbp ladder, lanes 1–4 represent the amplified products obtained at 55 °C, 58 °C, 60 °C and, 65 °C respectively. Although all exons were amplified at 65 °C, the maximum intensity for exon 20 was achieved at 58 °C (lane 2). In order to accommodate all the four exons in a single thermal cycler run, 58 °C was selected as the annealing temperature for the initial amplification for SEQ. Uncropped full images are given in Additional file 2: Figures S2-S4