In Situ Hybridization Method And Buffer

摘要

An improved method of in situ hybridization which relies on an improved formulation of the in situ hybridization buffer is described. In at least some formulations the buffer are non-toxic. The combination of Locked Nucleic Acid (LNA) comprising ISH probes and the improved ISH buffer are useful for detection of small non-coding RNA as well as in the manufacturing of ISH kits directed to the detection of such small non-coding RNA. Further disclosed is a method of semi-quantitative ISH and demonstration of the semi-quantitative ISHs diagnostic potential.

主权项

1. A formamide-free, detergent-free hybridization buffer for detecting small non-coding RNAs in formalin-fixed and paraffin-embedded (FFPE) tissue sections with LNA-probes by in situ hybridization, the buffer comprising:
a hybrid stabilizing agent comprising salts of mono-valent cations, salts of di-valent cations, or salts of mono-valent cations and di-valent cations; 0.5 to 5 M of a chaotropic component of urea, salts of guanidinium, salts of guanidine, or a mixture of two or more selected from the group consisting of urea, salts of guanidinium, and salts of guanidine; Denhardt's Solution; and a carrier RNA.