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Dramatically improved RNA in situ hybridization signals using LNA-modified probes.
- Source :
-
RNA (New York, N.Y.) [RNA] 2005 Nov; Vol. 11 (11), pp. 1745-8. Date of Electronic Publication: 2005 Sep 21. - Publication Year :
- 2005
-
Abstract
- 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.
- Subjects :
- DNA-Directed DNA Polymerase genetics
DNA-Directed DNA Polymerase metabolism
HSP70 Heat-Shock Proteins genetics
HSP70 Heat-Shock Proteins metabolism
Oligonucleotides
Oligonucleotides, Antisense chemistry
Poly A metabolism
Polynucleotide Adenylyltransferase genetics
Polynucleotide Adenylyltransferase metabolism
Saccharomyces cerevisiae genetics
Saccharomyces cerevisiae Proteins genetics
Saccharomyces cerevisiae Proteins metabolism
In Situ Hybridization, Fluorescence methods
Oligonucleotide Probes chemistry
Oligonucleotide Probes genetics
Oligonucleotide Probes metabolism
Oligonucleotides, Antisense genetics
Poly A chemistry
RNA, Fungal analysis
Subjects
Details
- Language :
- English
- ISSN :
- 1355-8382
- Volume :
- 11
- Issue :
- 11
- Database :
- MEDLINE
- Journal :
- RNA (New York, N.Y.)
- Publication Type :
- Academic Journal
- Accession number :
- 16177135
- Full Text :
- https://doi.org/10.1261/rna.2139705