Dr. Kees Fluiter, Amsterdam Medical Center:
“The Superior Probes of RiboTask are outperforming other probes for in situ applications to detect both miRNA and mRNA. These Superior Probes allow for a shortened in situ protocol that can be completed within 4 hrs. They can be used in a variety of applications including in cultured cells, paraffin embedded tissue and whole mount in situ hybridisations”
[B. S. Budde et al. Nature Genetics 2008, 40(9):1113-8; E. Aronica et al. Eur. J. Neurosci. 2010, 31(6):1100-7; Manuscripts in press]
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In situ hybridization is routinely used to visualize transcribed sequences in embryos, tissues, and cells. Classically, detection of mRNAs in tissues is done by hybridization with a labelled antisense RNA probe transcribed from a cDNA template. This method is relatively cumbersome and time consuming taking at least several days to perform.
Recently, locked nucleic acids (LNA) modified DNA probes were introduced with an increased affinity for a matching RNA sequence allowing the use of short probes which are resistent against nucleases. However, binding affinity of LNA-DNA probes is suboptimal as compared to the Superior Probes of RiboTask which are LNA-modified 2’-O-Me-RNA oligos. The Superior Probes display even stronger binding to complementary RNA while requiring 30% less LNA modifications as compared to LNA-DNA oligonculeotides.
The Superior Probes are only 19-22 mers in length and allow for an extremely efficient protocol for in situ hybridization detection of miRNA and mRNA that can be done in its entirety within 4 hours. These probes are suitable to detect alternatively spliced exons that are processed in a tissue specific manner. This new rapid protocol using superior probes will expand the general use of in situ hybridization for studies of transcriptional regulation and alternative splicing.
Contact RiboTask for further information and help on probe design.