Learn about the power of LNA (Locked Nucleic Acid) technology and QIAGEN's LNA enhanced product portfolio for RNA and DNA research. Download the slide deck!

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QIAGEN Locked Nucleic Acid (LNA®) Tools
Experience truly exceptional RNA research
The Power of LNA & QIAGEN‘s LNA Enhanced Portfolio

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Content
2Successfully Detect miRNAs Using qPCR with LNA Technology
The power of LNA
LNA enhanced portfolio
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LNA (Locked nucleic acids) are a class of high affinity
RNA analogs with the ribose ring locked in the ideal
conformation for Watson-Crick binding
• LNA increase the strength of base pairing
• LNA increase the binding affinity compared to
conventional DNA and RNA oligos
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LNA® - making oligos with unprecedented characteristics
• Increased Tm (2 - 8ºC per base)
• Improved mismatch discrimination
• High sensitivity and specificity in probes and assays
• Increased oligo stability and potency in cells
The power of LNA

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QIAGEN the new home for Locked Nucleic Acid
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Taking advantage of LNA requires sufficient experience:
o Knowledge of optimal LNA positioning in the oligo
o Understanding of RNA
o Understanding of the application and its purpose
EXIQON (a QIAGEN company) optimized application designs over many years
• Patented designs
• Sophisticated design algorithms behind every LNA-enhanced product
• 30-60 parameters in play when designing LNA oligo-based products:
• Tm, length, secondary structure/accessibility, mis-match discrimination/off-target binding,
binding specificity, self-complementarity, LNA -positioning
• Online design tools for custom products – now at QIAGEN
20 years
experience in
designing LNA
oligos
“Exiqon reagents stand out in front of
other platforms from a technological
standpoint”.
Dr Parker Antin, University of Arizona

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LNA is used to adjust the Tm of oligonucleotides
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Tm adjustments enables:
• Increased affinity towards DNA and RNA targets
• Increased specificity (shorter sequences)
• Increased flexibility in oligo design
• Sequence independence (GC content less important)
Same length, higher Tm Shorter length, similar Tm

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LNA enables detection of all microRNAs - even the AT-rich
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LNA ™
DNA
98% of these
microRNAs
have under
50% GC
66% of these
microRNAs
have under 50% GC
30% of these microRNAs
have under 50% GC
Increasing GC content

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LNA enables oligo Tm normalization
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IncreasingGCcontent
Power of Tm-normalization:
• Overall narrower Tm interval (red)
• Highly increased Tm of AT-rich assays
• Shift to higher median Tm (x-axis)
• Higher stringency and potency
Benefits of LNA & Tm-normalization
• Higher sensitivity and specificity
• Ability to address targets of low Tm e.g.
miRNA low in GC content
• Inhibitors: higher potency of all
inhibitors
• Detection probes: Uniform hybridization
of all miRNA at the same Thyb for ISH
(enables automation)
• qPCR: uniform amplification of all miRNA
targets

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LNA enhances the discriminatory power of oligonucleotides
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LNA enhances specificity of oligonucleotides for complementary RNA and DNA
targets
40% Form.
LNA DNA
THJ200
²rip 42 C 20‘
KD200
THJ202
THJ201
LNA DNA
50% Form.
B
40% Form.
LNA DNA
THJ200
²r
KD200
THJ202
THJ201
LNA
50% Form
B
mRNA SSA4 in situ hybridization,
fixed yeast cells.
Thomsen et al., 2005, RNA
Target
Probe
Perfect match
DNA
Single mismatch
3’ccaggaaggaaccac-5’
∆Tm
DNA 15mer
5’ggtccttacttggtg3’ Tm = 59.4°C Tm = 55.7°C 3.7°C
DNA/LNA 15mer
5’ggtccttActtggtg3’ Tm = 60.9°C Tm= 52.7°C 8.2°C

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LNA
DNA
RNA
OME
LNA ™ vs. competing technology
ISH using DIG-labeled probes (miR-122)
Discriminating miR-1 in heart of chicken embryo
Pictures kindly provided by D. Sweetman, University of East Anglia, Norwich, UK
Unique sensitivity Unique specificity
gga-miR-1a UGGAAUGUAAGGAAGUGUGUGG
gga-miR-206 UGGAAUGUAAAGAAGUAUGU
LNA enables RNA detection – the ONLY technology which works!
“Basically no other technology has been demonstrated to work for microRNA detection
in situ.”
Dr. Dylan Sweetman, University of East Anglia

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Exceptional specificity – single–nucleotide mismatch discrimination
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The signal obtained from perfectly matched LNA
oligos (100%) is compared to signal from LNA
oligos with a single nucleotide mismatch.
Perfect match
Single nucleotide mismatch

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Effective RNA silencing - with the robustness and stability for in vivo
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Content
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The power of LNA
LNA enhanced portfolio
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A solution for every step of your RNA workflow
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NGS PCR
Data
analysis
&
Interpret
ation
Function
al
analysis
Instruments
• QIAcube/QIAcube HT
• QIAsymphony® SP/AS
• QIAxcel Advanced
• QIAxpert (for QC)
• QIAxpert
• QIAxcel Advanced
• QIAgility
• Rotor-Gene® Q
• QIAsymphony SP/AS
Kits and Reagents
• RNeasy Plus Kits
• RNeasy Kits for all
sample types
• exoRNeasy Kits for
exosomal RNA
• PAXgene Blood/Tissue
RNA Systems for
stabilization and
purification
• Custom LNA®
Detection Probes for
mRNA and lncRNA
• QIAseq® UPX 3'
Transcriptome Kits
• QIAseq UPX 3' Targeted
RNA Panel
• QIAseq Stranded RNA
Library Kits
• QIAseq Targeted RNA
Panels
• QIAseq Targeted RNAscan
Panels
• QIAseq FX Single Cell RNA
Library Kit
• QIAseq Immune Repertoire
RNA Library Kits
• RT2 Profiler PCR Arrays
Predesigned assays and
pathway panels for
mRNA and lncRNA
• QuantiNova qPCR Kits
Combined with self-
designed primers or
QuantiTect® Primer
Assays
• Ingenuity® Pathway
Analysis (IPA®)
• CLC Genomics
Workbench
• Biomedical Genomics
Workbench
• GeneGlobe® Data
Analysis Center
• Antisense LNA
GapmeRs
For mRNA and lncRNA
silencing in vitro and in
vivo
• miRNeasy Kits for all
sample types
• PAXgene Blood/Tissue
miRNA Systems for
stabilization and
purification
• miRCURY LNA miRNA
Detection Probes
• QIAseq miRNA Library Kit
• QIAseq miRNA Library QC
PCR Panel and Assays
• miRCURY LNA miRNA
PCR System
Predesigned or custom
PCR assays and panels
• miRCURY LNA miRNA
Mimics, Inhibitors and
Target Site Blockers
For function analysis in
vitro and in vivo
QIAGEN Genomic Services: Sample preparation, NGS analysis, qPCR analysis, Data analysis and biological
interpretation
RNA
preparation
Localization
ISH
Northern
mRNAlncRNAmiRNA

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A solution for every step of your RNA workflow – LNA enhanced
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NGS PCR
Data
analysis
&
Interpret
ation
Function
al
analysis
Instruments
• QIAcube/QIAcube HT
• QIAsymphony® SP/AS
• QIAxcel Advanced
• QIAxpert (for QC)
• QIAxpert
• QIAxcel Advanced
• QIAgility
• Rotor-Gene® Q
• QIAsymphony SP/AS
Kits and Reagents
• RNeasy Plus Kits
• RNeasy Kits for all
sample types
• exoRNeasy Kits for
exosomal RNA
• PAXgene Blood/Tissue
RNA Systems for
stabilization and
purification
• Custom LNA®
Detection Probes for
mRNA and lncRNA
• QIAseq® UPX 3'
Transcriptome Kits
• QIAseq UPX 3' Targeted
RNA Panel
• QIAseq Stranded RNA
Library Kits
• QIAseq Targeted RNA
Panels
• QIAseq Targeted RNAscan
Panels
• QIAseq FX Single Cell RNA
Library Kit
• QIAseq Immune Repertoire
RNA Library Kits
• RT2 Profiler PCR Arrays
Predesigned assays and
pathway panels for
mRNA and lncRNA
• QuantiNova qPCR Kits
Combined with self-
designed primers or
QuantiTect® Primer
Assays
• Ingenuity® Pathway
Analysis (IPA®)
• CLC Genomics
Workbench
• Biomedical Genomics
Workbench
• GeneGlobe® Data
Analysis Center
• Antisense LNA
GapmeRs
For mRNA and lncRNA
silencing in vitro and in
vivo
• miRNeasy Kits for all
sample types
• PAXgene Blood/Tissue
miRNA Systems for
stabilization and
purification
• miRCURY LNA miRNA
Detection Probes
• QIAseq miRNA Library Kit
• QIAseq miRNA Library QC
PCR Panel and Assays
• miRCURY LNA miRNA
PCR System
Predesigned or custom
PCR assays and panels
• miRCURY LNA miRNA
Mimics, Inhibitors and
Target Site Blockers
For function analysis in
vitro and in vivo
QIAGEN Genomic Services: Sample preparation, NGS analysis, qPCR analysis, Data analysis and biological
interpretation
RNA
preparation
Localization
ISH
Northern
mRNAlncRNAmiRNA

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QIAseq® miRNA Library Kits
The QIAseq miRNA Library Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease 15
miRNA / piRNA sequencing, gel-free & LNA-enhanced
• Utilizes Unique Molecular Indices (UMI) technology
• No more gels – gel-free, bead-based workflow starting
with only 1 ng RNA
• For miRNA, piRNA and small RNA expression analysis
and novel small RNA discovery
• Data analysis included through cloud-based GeneGlobe
Data Analysis Site
QIAseq miRNA Library Kit (96), Cat. no. 331505
QIAseq miRNA Library Kit (12), Cat. no. 331502
QIAseq miRNA NGS 12 Index IL (12), Cat. no. 331592
QIAseq miRNA NGS 48 Index IL (96), Cat. no. 331595

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QIAseq miRNA Library QC PCR Panel and Assays
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For evaluating RNA sample quality and for assessing
NGS performance post-sequencing
• Unique qPCR-based sample QC of miRNA/small RNA
samples prior to NGS
• Essential for challenging samples with low RNA content,
such as biofluids
• LNA miRNA PCR Assays in ready-to-use PCR panels
• Compatible with all major qPCR instruments
• Comprehensive set of 52 RNA spike-ins, spanning a wide
range of concentrations
• Thorough post-sequencing assessment of NGS linearity
and reproducibility
QIAseq miRNA Library QC PCR Array Kit, Product no. 331541
QIAseq miRNA Library QC PCR Assay, Product no. 331551
QIAseq miRNA Library QC Spike-ins, Product no. 331535
The QIAseq UPX Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease

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QIAseq UPX 3' Transcriptome Kit
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High-throughput 3’-transcriptome NGS from ultralow
amounts of RNA
• Sample input from 1 to 100 cells or 10 pg to 1 ng isolated
RNA
• LNA -enhanced chemistry for increased accuracy,
specificity and sensitivity
• Integrated unique molecular indexing (UMI) removes
amplification bias
• Cell tagging and sample indexing enables simultaneous
sequencing of up to 18,432 transcriptomes
• Data analysis with the GeneGlobe Data Analysis Center
QIAseq UPX 3' Transcriptome Kit (96), Cat.no. 333088
QIAseq UPX 3' Transcriptome Kit (384), Cat.no. 333090
QIAseq UPX 3' Trans. 12-Index (48), Cat.no. 333074
QIAseq UPX 3' Trans. 48-Index (192), Cat.no. 333075
The QIAseq UPX Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease

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QIAseq UPX 3' Targeted RNA Panel
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High-throughput, targeted gene expression using
3’-targeted NGS
• Target up to 1000 genes using a cost-effective, time-
saving single-tube library prep
• LNA-enhanced chemistry for increased accuracy,
specificity and sensitivity
• Integrated unique molecular indexing (UMI) removes
amplification bias
• Cell tagging and sample indexing enables simultaneous
sequencing of up to 147,456 targeted libraries
• Cloud-based data analysis with the GeneGlobe Data
Analysis Center
QIAseq UPX 3' Targeted RNA Panel (96), Cat.no. 333041
QIAseq UPX 3' Targeted RNA Panel (384), Cat.no. 333043
QIAseq UPX 3' Targeted RNA 12 index (48), Cat.no. 333044
QIAseq UPX 3' Targeted 96 index A (384), Cat.no. 333051
QIAseq UPX 3' Targeted 96 index B (384), Cat.no. 333052
QIAseq UPX 3' Targeted 96 index C (384), Cat.no. 333053
QIAseq UPX 3' Targeted 96 index D (384), Cat.no. 333054
The QIAseq UPX Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease

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miRCURY LNA miRNA PCR System
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Unique miRNA qPCR system of universal RT combined with two miRNA-specific qPCR primers
Accurate and reliable quantification
• Accurate and reliable quantification of individual
miRNAs from as little as 1pg total RNA
Distinguish between miRNA sequences
• Distinguish between miRNA sequences that differ by a
single nucleotide
Thoroughly validated
• LNA-enhanced and Tm normalized primers,
Easy & Quick protocol
• 3-hour, easy-to-follow protocol that minimizes pipetting
Various control options
• Controls available for RNA isolation, RT and PCR
efficiency control
Data analysis
• GeneGlobe Data Analysis Center
One cDNA reaction for all miRNAs
Two LNA®-enhanced miRNA-
specific qPCR primers
The miRCURY LNA® miRNA PCR System is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a
disease

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miRCURY LNA miRNA PCR System
20Successfully Detect miRNAs Using qPCR with LNA Technology
Universal cDNA
synthesis
3 hour workflow
Individual or custom
Assays and
Ready-to-use panels
+
high performance miRNA
SYBR Green PCR Kit
Easy and free of
charge data
analysis
miRNA
isolation
cDNA
synthesis
PCR
Data
analysis
miRCURY LNA RT Kit miRCURY LNA miRNA PCR Assays
miRCURY LNA miRNA Custom PCR Assays
miRCURY LNA miRNA miRNome Panels
miRCURY LNA miRNA Focus Panels
miRCURY LNA miRNA QC Panels
miRCURY LNA SYBR Green PCR Kit
GeneGlobe Data Analysis
Center
Easy start with miRCURY LNA miRNA PCR Starter Kit + Optional RNA Spike-in Kit
for QC

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Antisense LNA GapmeRs
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Potent antisense oligonucleotides for highly efficient knockdown of mRNA and lncRNA
• Efficient RNase H dependent degradation of
complementary RNA targets
• Active in vivo and in vitro – enabling the
analysis RNA function in a wide range of
model systems
• Excellent alternative to siRNA for knockdown
of mRNA and lncRNA
• Taken up by cells by transfection or
unassisted delivery
• Designed with a sophisticated and
empirically developed algorithm for potent
and specific knockdown of target RNAs

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Antisense LNA GapmeRs
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Potent antisense oligonucleotides for highly efficient knockdown of mRNA and lncRNA
• 15 - 16mer LNA / DNA antisense oligonucleotide
• LNA at extremities enhances affinity for the target
• RNase H activity requires a certain distance (”gap”) between
LNAs
• Phosphorothioate modified backbone
• Online design tool
• Empirically derived design algorithm evaluates thousands of LNA
GapmeRs with more than 30 design parameters
Antisense LNA GapmeRs Standard, Product no. varies
Antisense LNA GapmeRs Custom Plate, Product no. 339530
The Antisense LNA GapmeRs is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease

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miRCURY LNA miRNA Inhibitors
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Inhibitors for miRNA loss-of-function studies
• Tm-normalized inhibitors with unmatched potency against
any miRNA, regardless of GC content
• Power Inhibitors so potent that they work by unassisted
uptake with no need for transfection reagents
• Superior specificity and biological stability for long-lasting
antisense activity
• Available in 1-, 5- and 15 nmol quantities
• Fluorescent labels for convenient monitoring of
transfection efficiency
miRCURY LNA miRNA Inhibitors, Product no. varies
miRCURY LNA miRNA Power Inhibitors, Product no. varies
Custom miRCURY LNA miRNA Inhibitors, Product no. varies
miRCURY LNA miRNA Family Power Inhibitors, Product no. varies
miRCURY LNA miRNA Inhibitor Libraries, Product no. varies
miRCURY LNA miRNA Inhibitors are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of
a disease

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miRCURY LNA miRNA Mimics
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For studies on miRNA function and gene regulation using synthetic miRNA
• Highly potent, mature miRNA mimics with
unique triple RNA strand design
• No miRNA-star activity from bisected LNA
-enhanced passenger strand
• miRNA strand sequence matches
miRBase annotation
• Available with fluorescent label to assess
transfection efficiency
• Available with biotinylated miRNA strand
to isolate targets by RNA pull-down
miRCURY LNA miRNA Mimic (5 nmol) / (20 nmol),
Product no. 339173 / 339174
miRCURY LNA Premium miRNA Mimic (5 nmol) /
(20 nmol), Product no. 339178 / 339179
miRCURY LNA miRNA Mimics are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a
disease

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miRCURY LNA miRNA Power Target Site Blockers
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For studying the effects of an individual miRNA on a single target site
• Custom-designed target site blockers for specific inhibition of
miRNA targets
• Sophisticated design and superior high affinity, regardless of
target sequence
• Unmatched high efficacy in vitro and in vivo
• Unrivaled performance and high protein expression due to
lack of RNase H-dependent mRNA degradation
• Efficient at very low concentrations, outcompeting miRNAs for
their target sites
• Superior biological stability for long-lasting antisense activity
miRCURY LNA miRNA Power Target Site Blockers (5 nmol) / (15 nmol),
Product no. 339194 / 339195
miRCURY LNA miRNA Power Target Site Blockers, in vivo Ready (5 nmol) /
(15 nmol), Product no. 339199 / 339200
TSB
miRCURY LNA miRNA Power Target Site Blockers are intended for molecular biology applications. These products are not intended for the diagnosis,
prevention, or treatment of a disease

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miRCURY LNA miRNA Detection Probes
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For ultra-sensitive & specific miRNA detection by in situ hybridization (ISH) or Northern blotting
• Superior sensitivity and specificity for detecting low-abundance
miRNAs
• Predesigned probes available for all miRNA species or
Custom-designed probes for targeting novel miRNA
sequences
• A wide selection of available labels enables multiplexing and
co-localization
• Ideal for standardized protocols for automated, high-
throughput ISH
• Fast and easy workflow using the One-day miRNA ISH
protocol
miRCURY LNA miRNA ISH Buffer Set (FFPE), Product no. 339450
miRCURY LNA miRNA ISH Optimization Kit (FFPE) 1-9, Product no. varies
miRCURY LNA miRNA ISH Buffer and Controls, Product no. 339459
miRCURY LNA miRNA ISH Optimization Kits (FFPE) are intended for molecular biology applications. These products are not intended for the diagnosis,
prevention, or treatment of a disease

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Custom LNA mRNA Detection Probes
miRCURY LNA miRNA ISH Optimization Kits (FFPE) are intended for molecular biology applications. These products are not intended for the diagnosis,
prevention, or treatment of a disease
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For ultra-sensitive detection and discrimination by highly similar mRNA and lncRNA
by in situ hybridization and Northern blotting
• Sensitivity and specificity superior to DNA probes and
riboprobes
• Short probes ideal for discriminating highly similar sequences -
e.g. splice variants and isoforms
• Online tools provide optimally designed LNA -enhanced
probes in minutes
• Available with a wide selection of labels
• No cloning expertise required
• Excellent tissue penetration

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Custom LNA ™ Oligonucleotides
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For experiments requiring custom-designed, LNA -enhanced oligonucleotides
QIAGEN offers synthesis of custom LNA oligonucleotides with a
wide variety of modifications, labels, synthesis scales, purification
scale options
• Create or improve any RNA or DNA application with LNA –
ONLY with QIAGEN
• Order any LNA -enhanced oligo of interest
• your own design with the LNA spiking pattern of your choice
• Use the online LNA oligo help tools to for Tm prediction and
LNA optimization
• LNA oligos are available in small scale (nmol-µmol) for in vitro
applications and in large scale (mg) for in vivo use in animal
models
Alternatively, have QIAGEN experts in LNA oligo design assist in
designing the most optimal LNA oligo for their application (design
fee applicable)
Custom LNA mRNA Detection Probes are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or
treatment of a disease.

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Website link
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For detailed information about LNA products please visit us: www.qiagen.com/LNA