Dramatically improved RNA in situ hybridization signals using LNA-modified probes | Zendy

Rune Thomsen | Zendy; Peter Stein Nielsen | Zendy; Torben Heick Jensen | Zendy

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Dramatically improved RNA in situ hybridization signals using LNA-modified probes

Author(s) -

Rune Thomsen,

Peter Stein Nielsen,

Torben Heick Jensen

Publication year - 2005

Publication title -

rna

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 3.037

H-Index - 171

eISSN - 1469-9001

pISSN - 1355-8382

DOI - 10.1261/rna.2139705

Subject(s) - locked nucleic acid , biology , oligonucleotide , rna , nucleolus , in situ hybridization , nucleic acid , dna , microbiology and biotechnology , hybridization probe , in situ , molecular probe , nucleic acid thermodynamics , complementary sequences , oligomer restriction , computational biology , messenger rna , biochemistry , nucleus , gene , chemistry , statistics , mathematics , organic chemistry

In situ detection of RNA by hybridization with complementary probes is a powerful technique. Probe design is a critical parameter in successful target detection. We have evaluated the efficiency of fluorescent DNA oligonucleotides modified to contain locked nucleic acid (LNA) residues. This increases the thermal stability of hybrids formed with RNA. The LNA-based probes detect specific RNAs in fixed yeast cells with an efficiency far better than conventional DNA oligonucleotide probes of the same sequence. Using this probe design, we were also able to detect poly(A)(+) RNA accumulation within the nucleus/ nucleolus of wild-type cells. LNA-based probes should be readily applicable to a diverse array of cells and tissue samples.

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