Technical Documents
Description
Locked Nucleic Acid (LNA) was first described by Wengel and co-workers in 19981 as a novel class of conformationally restricted oligonucleotide analogues. LNA is a bicyclic nucleic acid where a ribonucleoside is linked between the 2’-oxygen and the 4’-carbon atoms with a methylene unit. Oligonucleotides containing LNA exhibit unprecedented thermal stabilities towards complementary DNA and RNA2, which allows excellent mismatch discrimination. In fact the high binding affinity of LNA oligos allows for the use of short probes in, for example, SNP genotyping3, allele specific PCR and mRNA sample preparation. In fact, LNA is recommended for use in any hybridization assay that requires high specificity and/or reproducibility, e.g., dual labelled probes, in situ hybridization probes, molecular beacons and PCR primers. Furthermore, LNA offers the possibility to adjust Tm values of primers and probes in multiplex assays. As a result of these significant characteristics, the use of LNA-modified oligos in antisense drug development is now coming under investigation4, and recently the therapeutic potential of LNA has been reviewed.5 LNA can be mixed with DNA and RNA, as well as other nucleic acid analogues, modifiers and labels. LNA oligonucleotides are water soluble, and can be separated by gel electrophoresis and precipitated by ethanol.
Details
Usage
- Coupling: 3 minute coupling time recommended. Increase oxidation time 3 fold for optimal performance.
- Deprotection: No changes needed from standard method recommended by synthesizer manufacturer.
Specifications | |
---|---|
Diluent | Anhydrous Acetonitrile |
Storage | Freezer storage, -10 to -30�C, dry |
Stability | 2-3 days |
Dilution/Coupling Data
The table below show pack size data and, for solutions, dilution and approximate coupling based on normal priming procedures.
ABI 392/394
Catalog # | Pack Size | Grams/Pack | 0.1M Dil. (mL) | Approximate Number of Additions | |||||
---|---|---|---|---|---|---|---|---|---|
LV40 | LV200 | 40nm | 0.2μm | 1μm | 10μm | ||||
10-2030-05 | 0.5 g | 0.5 | 6.47 | 202.33 | 121.4 | 75.88 | 55.18 | 40.47 | 10.12 |
10-2030-10 | 1.0 g | 1 | 12.94 | 418 | 250.8 | 156.75 | 114 | 83.6 | 20.9 |
Expedite
Catalog # | Pack Size | Grams/Pack | Dilution (mL) | Approximate Number of Additions | ||||
---|---|---|---|---|---|---|---|---|
Molarity | 50nm | 0.2μm | 1μm | 15μm | ||||
10-2030-05 | 0.5 g | 0.5 | 9.66 | 0.07 | 186.8 | 116.75 | 84.91 | 11.68 |
10-2030-10 | 1.0 g | 1 | 19.31 | 0.07 | 379.8 | 237.38 | 172.64 | 23.74 |
References
(1a)A.A. Koshkin, S.K. Singh, P. Nielsen, V.K. Rajwanshi, R. Kumar, M. Meldgaard, C.E. Olsen, and J. Wengel, Tetrahedron,1998, 54, 3607-3630.(1b) S.K. Singh, P. Nielsen, A.A. Koshkin, and J. Wengel, Chem. Comm., 1998, (4), 455-456. (2) L. Kværnø and J. Wengel, Chem. Comm., 1999, (7), 657-658.(3) P. Mouritzen, A.T. Nielsen, H.M. Pfundheller, Y. Choleva, L. Kongsbak, and S. Møller, Expert Review of Molecular Diagnostics, 2003, 3(1), 27-38.(4a)J. Kurreck, E. Wyszko, C. Gillen, and V.A. Erdmann, Nucleic Acids Res., 2002, 30, 1911-1918. (4b)H. Ørum and J. Wengel, Curr. Opinion in Mol. Therap., 2001, 3, 239-243.(5a)M. Petersen and J. Wengel, Trends in Biotechnology, 2003, 21(2), 74-81. (5b)D.A. Braasch, D.R. Corey, Biochemistry, 2002, 41, 4503-4510.