Table 2.
Details of PCR amplification of a genomic DNA template.
| Symbol | size (bp) |
Tm (°C) |
Cq (cycles) |
|---|---|---|---|
| 18SrRNA | ~170 | 84.3 | 14.8 |
| Arm | no amplification | no amplification | no amplification |
| EF1a# | light smear | 3 peaks: 76.6; 79.8; 83.7 | 35.8 |
| RpL32 | ~100 | 78.4 | 28.8 |
| GAPDH# | light smear | 79.8 | 34.9 |
| Actin | ~650 | no amplification | no amplification |
| SDHa | no amplification | no amplification | no amplification |
| AnnIX | light smear | no amplification | no amplification |
Genomic DNA amplicons were sized on an agarose gel following 30 cycles of PCR amplification using a Taq polymerase from Qiagen, a melting temperature of 58°C, and 50 ng of template. The melting temperatures (Tm) and Cq values were determined by quantitative PCR on a LightCycler 480 Instrument (Roche) as detailed in the main text and on 5 ng of genomic DNA. # means that primers span an exon-exon boundary. Alignments of complementary and genomic DNA sequences can be found for Actin, GAPDH, and EF1a in Additional file 3.