Before revision: Although introducing base pair mismatch in some sets of the probes could minimize annealing temperature difference between each sets of probes, the hybridization problem is still significant if more than 10-20 targets are tested in one reaction.
After revision: Although introducing base pair mismatch in some sets of the probes could minimize annealing temperature differences between each set of probes, the hybridization problem is still significant if more than 10-20 targets are tested in one reaction.