Detection of immobilized nucleic acid

A method for determining the presence or absence of nucleic acid in a sample, wherein the method comprises;

• a) combining an unsymmetrical cyanine dye compound characterized as being essentially non-genotoxic with a sample to prepare a labeling mixture, wherein the sample is immobilized on a solid or semi-solid support;
• b) incubating the labeling mixture for a sufficient amount of time for the dye to associate with the nucleic acid to prepare an incubated sample;
• c) illuminating the incubated sample with an appropriate wavelength to prepare an illuminated sample;
• and
• d) observing the illuminated sample whereby the presence or absence of the nucleic acid is determined;
• with the proviso that the cyanine dye compound is not thiazole orange when the support is a polymeric gel.

A staining solution for determining the presence or absence of immobilized nucleic acid on a solid or semi-solid support, wherein the solution comprises:

• e) an unsymmetrical cyanine dye compound having formula
• wherein X is O, S or C(CH3)2;
• R1 is a fused benzene, methoxy, a C1-C6 alkyl;
• R2 and R3 are independently a C1-C6 alkyl;
• R4 is a C1-C6 alkyl or a methoxy;
• t is independently 0, 1, 2, 3, or 4;
• s is independently 0, 1, 2, 3 or 4 and
• n is 0, 1, 2 or 3;
• and
• a) a tris borate or tris acetate buffer;
• with the proviso that the cyanine dye compound is not thiazole orange.

A kit for the detection of immobilized nucleic acid, the kit comprising:

• f) a compound having the formula
• wherein X is O, S or C(CH3)2;
• R1 is a fused benzene, methoxy, a C1-C6 alkyl;
• R2 and R3 are independently a C1-C6 alkyl;
• R4 is a C1-C6 alkyl or a methoxy;
• t is independently 0, 1, 2, 3, or 4;
• s is independently 0, 1, 2, 3 or 4 and
• n is 0, 1, 2 or 3;
• with the proviso that the cyanine dye compound is not thiazole orange.