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 Product Sheet
LNA Oligonucleotides exhibit unprecedented thermal stabilities towards complementary DNA and RNA.
Excellent mismatch discrimination.
The high binding affinity of LNA oligos allows for the use of short probes & hybridization assays.
LNA offers the possibility to adjust Tm values of primers and probes in multiplex assays.
Use in antisense protocols, hybridization assay, in situ hybridization probes, dual labeled probes, molecular beacons and PCR primers.
LNA bases incorporated in oligonucleotides show remarkable hybridization specificity to both RNA and DNA. The higher Tm from 2-4oC of each modified LNA base offers excellent duplex stability. LNA-C ('C') and 5-methyl-C ('mC') are equivalent in terms of duplex stability.
LNA containing oligonucleotides are substrates for most enzymes commonly used in molecular biology. A selection is listed below:
3’ LNA is substrate for polymerases
LNA primers and as template are read by different polymerases (Klenow, Taq polymerase) and reverse transcriptase.
LNA can be cut by restriction enzymes
32P labeling with T4 polynucleotide kinase
3’ LNA enhances resistance to exonuclease I
The following guideline represents the wide application of LNA use for almost all oligo design and use. Gene Link technical service offers advice on oligo design and use of modifications based on application.
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Application
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Recomended Modifications
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Antisense Gene Target
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Oligonucleotides containing 2'-OMe-nucleotides (2'-OMe-RNA) forms more stable hybrids with complementary RNA strands than equivalent DNA and RNA sequences.
Phosphorothioate linkages confer to the oligonucleotides resistance to nuclease degradation
Locked Nucleic Acids (LNA) has demonstrated an unsurpassed duplex stability. Use phosphorothioate linkages to impart nuclease resistance and LNA bases to achieve most stable hybridization and thus duplex stability.
LNA-DNA-LNA gapmers. Substitue 4 LNA bases at the ends with 8-10 DNA bases in
the middle. This activates RNase H activity. These are ~10 times more stable
than siRNA but ~6 fold less knockout is achieved.
3’ Cholesterol modification helps in cellular uptake. |
| RNA Interference (siRNA) |
LNA substituted bases at the 3’ and 5’ end of siRNA enhances duplex
stability and increases exonuclease resistance. It has been shown that siRNA
with end modified LNA bases have ~10 fold longer half life. [Olaf et al. Mol
Cancer Ther 2007; 6(3):833–43].
3-4 LNA base substitution in the sense strand increases duplex stability.
2’F U and C substituted siRNA are more resistant to RNAse degradation.
3’ Cholesterol modification helps in cellular uptake.
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Real-Time PCR probes, QPCR
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LNA substituted bases in the probe enhances the duplex stability and thus shorter probes can be synthesized.
The use of LNA bases renders the probe greaterduplex stability than the use of single MGB (minor grove binders) at the 3’ end. It is an excellent substitute for TaqMan MGB probes.All types of fluorescent dyes and backbone modifications can be performed.
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SNP Genotyping, Allelic Discrimination.
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LNA substituted bases imparts greater specificity with higher Tm.
All types of fluorescent dyes and backbone modifications can be performed.
C-5 methylated pyrimidine deoxy-nucleosides behave similar to LNA bases in imparting duplex stability.
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Hybridization Probes and PCR Amplification Primers.
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LNA substituted bases imparts greater specificity with higher Tm. Substitute 4-6 bases with LNA . |
*License Agreement: Locked-nucleic Acid (LNA) phosphoramidites are protected by EP Pat No. 1013661, US Pat No. 6,268,490 and foreign applications and patents owned by Exiqon A/S. Products are made and sold under a license from Exiqon A/S. Products are for research purposes only. Products may not be used for diagnostic, clinical, commercial or other use, including use in humans. There is no implied license for commercial use, including contract research, with respect to the products and a license must be obtained directly from Exiqon A/S for such use.
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