If the cycle annealing
temperature is below the Tm of a primer set, then they may start to anneal in places of less than perfect homology, leading to spurious false-positive amplicon production.
The program for N1S1 PCR assay includes initial denaturation (95[degrees]C x 10'), cycle denaturation (94[degrees]C x 1.30'), cycle annealing
(60[degrees]C x 1.30'), cycle extension (72[degrees]C x 1.50') and final extension (72[degrees]C x 30') with 35 numbers of cycles.
Figure 7 shows the changes in hole depth and hole area during cycle annealing
. It can be deduced from these uniform curves with various curing conditions that the curing condition does not affect the temperature dependence behavior, mainl struciural relaxation, of the epoxy resin.