The effects of probes with dInosine or wobbles in the same positions were also investigated in 3M TMAC. The presence of a dInosine in a specific position instead of a wobble would theoretically decrease the degeneration of the probe and subsequently increase the concentration of the particular probe variant. A probe with 21 N wobbles, wobbN_21, at the same positions as the dInosines in the Ino21 probe, was tested. The surprising result was that the probe containing N wobbles hybridized very well with the InflA target (29% of the total MFI, 2220 MFI, Figure 2C ) and the 21-pm target (29% of the total MFI, 2190 MFI, Figure 2C ). This is in the same range as hybridization of the Ino21 probe with the InflA (26% of the total MFI, 1976 MFI, Figure 2C ) and 21-pm (11% of the total MFI, 794 MFI, Figure 2C ) targets (39 and 37% of the total MFI versus the InflA and the 21-pm targets, see Figure 2A ). These results also demonstrate that the wobb_N21 probe is not affected to the same extent as the Ino21 probe by increasing the hybridization temperature from 45 to 55 C ( Figure 2C ). The test was repeated by comparing an Ino18 with a wobbN_18 probe ( Figure 3C and D) . At 45 C, the Ino18 probe hybridized at least as well as the wobbN_18 probe while, at 55 C, the wobbN_18 probe hybridized better than the Ino18 probe. Interestingly, a probe containing 24 wobbles still hybridized better with all 33%xnt and 33%xF targets (30-45% of the total MFI at 45 C; 13-22% of total MFI at 55 C; Figure 3C ) compared with Ino24 (6-16% of the total MFI at 45 C; 1% of the total MFI at 55 C; Figure 3A and B). Obviously, the 70-mer probes can accommodate multiple degenerate positions and still hybridize because the majority of probe molecules will contain several long perfectly matching stretches created by chance. This is further deliberated under Discussion section.