Figure 2
RT-qPCR optimisation of the NBS1 gene. (A) Amplification curves for the nine primer concentration combinations of the primer optimisation matrix. No amplification was seen for the no template controls (NTCs). The primer concentration combinations are indicated in the same colour as the corresponding curves. The mean Cq values and standard deviations (STD) were calculated from the amplification curves as outlined in Material and Methods. The primer concentration combinations are given in nmol/L for the forward and the reverse primer, respectively. (B) Digital image of NBS1 RT-qPCR reactions obtained for the primer optimisation matrix. Quality control of the RT-qPCR reactions by gel electrophoresis revealed only the NBS1 amplicon without additional, non-specific bands. The sizes of the molecular weight markers (MW) are given on the left, whereas the size of the NBS1 amplicon is indicated on the right (arrow). Wells loaded with RT-qPCR reactions are labelled with (+), while the respective no template control is labelled with (-). Sizes are given in base pairs and primer concentration combinations are given in nmol/L respectively.