abstract |
In an assay for a target nucleotide sequence employing hybridization with a target probe nucleotide sequence, cross reaction between the target probe and a non-target nucleotide sequence, capable of hybridizing with the target probe, is prevented or diminished by introducing, at some point during the hybridization, a non-target probe which hybridizes preferentially with the non-target nucleotide sequence. The melting temperature for the non-target probe/non-target nucleotide sequence hybrid is preferably greater than that of the target probe/non-target sequence hybrid. The hybridization can be carried out under isothermal conditions or preferentially with gradual cooling. |