Composition and methods for hybridization
The present invention relates to a composition for use in hybridization, the composition comprising: (a) at least one hybridization probe (“component (a)”); (b) at least one polar protic or polar aprotic solvent (“component (b)”) having a cyclic molecule structure; and (c) at least one carboxylic acid amide and/or salts thereof (“component (c)”), in an amount of more than 10 vol %, based on the composition. The invention furthermore relates to a use of this composition, and to methods for detecting nucleic acids and/or chromosomal aberrations in a biological sample by way of hybridization, using the composition.
1 . In a composition for detecting nucleic acids and/or chromosomal aberrations in a biological sample via in situ hybridization, the composition comprising at least one fluorescent-labeled locus-specific hybridization probe (“component (a)”) and water as a carrier or excipient, wherein the hybridization probe is selected from DNA, RNA, and locked nucleic acids (LNA), wherein the improvement comprises:
that the composition comprises:
2-pyrrolidone or 2-piperidone (“component (b)”) in an amount of 2 to 13 vol % or wt %, based on the composition, and
formamide and/or salts thereof (“component (c)”) in an amount of 20 to 30 vol % based on the composition, wherein the composition comprises said component (b) and said component (c) in a volume-based ratio in the range of 1:10 to 1:3, said composition further comprising:
dextran sulfate (“component (d)”), and
at least one inorganic salt (“component (g)”) wherein the inorganic salt is sodium chloride.
2 . The composition according to claim 1 , wherein the composition comprises said dextran sulfate in an amount in the range of 10 to 20 wt %, based on the composition.
3 . The composition according to claim 1 , wherein the composition comprises said inorganic salt in an amount in the range of 0.5 to 5 wt %, based on the composition.
4 . The composition according to claim 1 , said composition further comprising at least one chemical buffer system (“component (e)”).
5 . The composition according to claim 4 , where the at least one chemical buffer system is a citrate-based buffer system in an amount in the range of 0.1 to 3 wt %.
6 . The composition of claim 5 , wherein said citrate-based buffer system is an SSC (saline-sodium citrate) buffer system.
7 . The composition according to claim 1 , said composition further comprising at least one blocking and/or stabilizing agent (“component (f)”) in an amount in the range of 0.1 μg/μl to 10 μg/μl.
8 . The composition according to claim 1 , further comprising at least one detergent and/or surfactant.