Introducing a dInosine in every third position throughout the Ino24 probe decreased the hybridization dramatically for all the 33% targets (6-16% of the total MFI, 361-920 MFI at 45 C; 1% of the total MFI, 53-79 MFI at 55 C). When utilizing the 21Ino_9nt5 0 probe with the 33%xnt targets, the MFI decreased because of the mismatches created by the two sm at the 3 0 end of the target (4-10% of the total MFI, 232-415 MFI at 45 C). In comparison, the 21Ino_9nt5 0 probe and the 33%xF targets, with no mismatch, hybridized at 26-43% of the total MFI (1692-2898 MFI at 45 C). Furthermore, the 21Ino_9nt3 0 , which did cover the two sm at the 3 0 end of 33%xnt, hybridized more strongly (10-21% of total the MFI, 450-695 MFI at 45 C) than the 21Ino_9nt5 0 probe (4-10% of the total MFI, 232-415 MFI at 45 C). Both these results demonstrate that using the less stringent temperature (45 C) permits hybridization even when a large number of Inosines is present (21 dInosines in a 70-mer probe), as long as a matching region of at least 9 nt is formed in the hybrid.