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Experience truly exceptional RNA research with QIAGEN's next-generation, LNA®-enhanced tools. LNA (Locked Nucleic Acid) oligos bind with much higher affinity and specificity to RNA targets than standard DNA and RNA oligos – This enables specific and sensitive detection of small RNAs and discrimination between highly similar
sequences.
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Take your RNA research to the next level with QIAGEN LNA tools!
1. Take your RNA
research to the next
level with QIAGEN LNA tools
Experience truly exceptional RNA research with our next-generation, LNA®
-enhanced tools. LNA (locked
nucleic acids) oligos bind with much higher affinity and specificity to RNA targets than standard DNA and RNA
oligos – This enables specific and sensitive detection of small RNAs and discrimination between highly similar
sequences. We have merged QIAGEN’s cutting edge technologies with Exiqon’s 20 years of experience in LNA
oligo design to enhance your qPCR assays, in situ hybridization and functional analysis in cells and animals,
miRNA and RNA sequencing and much more!
Sample to Insight
2. 2 Take your RNA research to the next level with QIAGEN LNA tools 05/2018
Figure 2. LNA Tm
normalization. The affinity of traditional, full
length, microRNA inhibitors without LNA is highly influenced
by GC-content, with Tm
values spanning >40˚C. The Tm
of
miRCURY LNA microRNA inhibitors span just 10˚C around
an optimal high temperature, ensuring uniform, higher
potency.
30
40
50
RNA Tm
miRNA GC content (%)
60
70
80
Traditional DNA inhibitors LNA inhibitors
• Exceptional sensitivity: Detect RNA expressed at very low levels or in complex
samples i.e. liquid biopsies
• Superior specificity: Accurately discriminate between closely related sequences –
Achieve even single-nucleotide mismatch discrimination
• Higher affinity: Design more potent and sensitive oligos. Enables the design
of shorter probes or primers to target specific isoforms and helps overcome the
difficulties in studying very short sequences i.e. microRNAs
• Enhanced thermal stability: Achieve uniform detection regardless of GC content
with the freedom to place probes and primers where needed
• Resistant to exo- and endonucleases: Benefit from long lasting stability in vitro
and in vivo
LNA (Locked Nucleic Acids) are a
class of high-affinity RNA analogs in
which the ribose ring is “locked” in
a sterical conformation which has the
unique property of greatly enhancing the
strength of Watson-Crick base-pairing.
The power of LNA technology
LNA substitutions increase oligo melting temperature, enabling Tm
adjustment and normalization, regardless of oligo length or
GC content. LNA is optimal for designing short, high-affinity PCR primers, detection probes and antisense oligonucleotides.
Figure 1. Substituting LNA
for DNA gives higher
melting temperatures.
Unmatched hybridization affinity - regardless of GC content
3. Take your RNA research to the next level with QIAGEN LNA tools 05/2018 3
LNA increases the Tm
difference between perfectly matched and mismatched targets – giving an increased signal-to-noise
ratio. This broadens the detection window and boots assay specificity. Even closely related sequences that differ by as little
as one nucleotide can be accurately discriminated, in ISH studies, qPCR profiling and silencing.
Figure 4. Superior signal-to-noise ratios. The signal obtained
from perfectly matched LNA oligos (100%) is compared to
signal from LNA oligos with a single nucleotide mismatch.
Exceptional specificity – achieve single–nucleotide mismatch
discrimination
Due to their short length, high-affinity, LNA antisense oligonucleotides are taken up naturally by cells. Highly potent once
inside cells and resistant to enzymatic degradation, LNA antisense oligos effectively silence RNA in a broad range of animal
tissues – making them the tool of choice for revealing RNA functions and evaluating their therapeutic potential.
Figure 5. Efficient in vivo knockdown with LNA GapmeRs in
a broad range of tissues. The Antisense LNA GapmeR for
knockdown of Malat 1 in mice was injected subcutaneously
over a period of 5 weeks. Tissue samples from the mice were
collected 2 days after the final LNA GapmeR administration.
The results show highly efficient knockdown of Malat1 in a
broad range of different tissues.
MALAT1 RNA level %
(relative to PBS control)
120
100
80
60
40
20
0
2 days after last dose of MALAT1 LNA GapmeR
Liver
Jejunum
Ovaries
Kidney
Heart
Fatpad
Skeletal
muscle
Skin
Spleen
Lung
Brain
Effective RNA silencing - with the robustness and stability for animal
models
Figure 3. Ultra-sensitive and specific miRNA detection with LNA. Detection of SSA4 RNA in
∆rip1 fixed yeast cells using a single-labeled Cy3 Custom LNA mRNA Detection Probe (left)
or a single-labeled Cy3 DNA oligonucleotide (right) of the same length.
LNA DNA
THJ202
LNA inclusion in oligos vastly increases binding affinity and allows the
design of highly sensitive qPCR and ISH detection assays. Detect more
transcripts – even miRNA and lncRNA at low expression levels. Use
less sample material or complex samples with low RNA content.
Unsurpassed sensitivity – see the difference LNA technology makes