For in-depth pathway- or disease-specific gene expression analysis using LNA-enhanced, probe-based PCR

Features

Expert-designed panels targeting the most relevant pathway- or disease-focused genes

Simultaneous profiling of 84 related genes

Exceptional sensitivity and specificity using short, LNA-enhanced primers and FAM-labeled hydrolysis probes

Stability for room-temperature setup allowing automation

Robust and reproducible results in less than 2 hours using the QuantiNova Probe PCR Kit

Product Details

QuantiNova LNA Probe PCR Focus Panels enable quick and reliable gene expression analysis of over 150 pathways each for human, mouse or rat using hydrolysis probe-based qPCR. LNA enhancement enables us to design shorter primers with optimal Tm, allowing flexible positioning on the target for optimal sequence discrimination. Thorough design validation ensures top performance and robust detection. Optimized using QuantiNova chemistry, the simple workflow can be completed in under 2 hours and is robust enough to support room-temperature setup, automation and delayed run. We also offer Flexible Panels, which are semi-customizable from the assays in our Focus Panels, as well as Custom Panels with your choice of predesigned and/or custom assays.

Performance

Expert LNA enhancement for top qPCR performance

The assays in the QuantiNova LNA Probe PCR Focus Panels have been developed using stringent design criteria and lab-validated algorithms. Twenty years of LNA design experience has enabled us to optimize our sophisticated LNA design algorithm, which incorporates over 50 different parameters to guarantee the most optimal assay for successful target detection. Each primer–probe set delivers the highest specificity and efficiency for the most reliable and accurate results. LNA enhancement enables Tm normalization across the panel and gives the primers and probes higher binding affinity, dramatically increasing assay sensitivity and specificity.

The increased sensitivity ensures excellent amplification efficiencies down to 1 RNA molecule, making it easier for you to detect low-abundance targets such as lncRNAs from less starting material. Increased specificity from the clever placement of LNA gives you a high signal-to-noise ratio, allowing discrimination of sequences that differ by only a single nucleotide and eliminating non-specific amplification and primer-dimer formation.

The high binding affinity of LNA bases increases the flexibility of primer and probe placement on the transcript, so we are able to use intelligent positioning to design assays for otherwise difficult-to-analyze targets. This gives you robust and reliable quantification, even for AU-rich targets, low-abundance transcripts, targets with high secondary structure and highly complex samples.

Principle

Robust and high-performance hydrolysis probe-based qPCR

QuantiNova LNA Probe PCR Focus Panels use FAM-labeled hydrolysis probe-based detection, which is highly sensitive and robust, because only the desired PCR product is detected. Probe-based detection still requires high PCR specificity to prevent amplification artifacts such as non-specific PCR products and primer–dimers that can compete for reaction components and compromise performance. However, we’ve already optimized the QuantiNova LNA Probe PCR system for you to eliminate non-specific amplification and give you accurate and sensitive detection every time.

Most comprehensive and specific coverage

QuantiNova LNA Probe PCR Focus Panels include the assays needed for a thoroughly researched panel of relevant pathway- or disease-focused genes. The predesigned assays have been designed using our proprietary algorithm to provide the most sensitive, accurate and effective analysis. Most of the assays are intron-spanning when possible and detect only RNA. Assays that do not span an intron are designated as such, and if there is one exon in the target, unwanted signals can be easily eliminated using the QuantiNova Reverse Transcription Kit with the integrated gDNA removal step.

Flexible layout and controls

QuantiNova LNA Probe PCR Focus Panels are available in 96-well plate, 384-well plate and 100-well disc formats and are used to monitor the expression of 84 genes related to a disease state or pathway, plus multiple reference genes. Each Focus Panel includes control elements for the following:

Data normalization

Genomic DNA contamination detection

RNA sample quality

General PCR performance

Multiple plate discounts

Plate quantity ordered Discount (%)

1–5                                       0

6–11                              25

12–23                              40

24+                                      55

Complimentary data analysis

The complimentary, web-based data analysis tool in the GeneGlobe Data Analysis Center includes a user-friendly wizard to guide you step-by-step through the normalization and analysis of your data and generates publication-ready results figures.

Procedure

Choose one- or two-step qRT-PCR

QuantiNova LNA Probe PCR Focus Panels have been optimized using the time-tested and high-performance QuantiNova chemistry. Best results are obtained when reverse transcription is performed with the QuantiNova Reverse Transcription Kit. The resulting cDNA is then quantified by qPCR using the master mixes of the QuantiNova Probe PCR Kit combined with the assays on your panel.

For one-step qRT-PCR, we recommend using the QuantiNova Probe RT-PCR Kit, which offers quick reaction setup without need for optimization of reaction conditions. Simply combine RNA template with the ready-to-use RT-PCR master mix and the assays on your panel and start the reaction.

Fast and easy workflow

The fast and easy workflow can be completed in under 2 hours with minimal hands-on time and can be automated to save even more time and labor. The simple protocol has been tested for compatibility on all major brand qPCR cyclers . Furthermore, the cDNA generated in the RT step can be used across the entire QuantiNova LNA Probe PCR System, allowing seamless transition from assays to panels depending on your research needs, saving you time and sample.

What you need to get started with the QuantiNova LNA Probe PCR system

Reverse transcription: QuantiNova Reverse Transcription Kit

qPCR mastermix: QuantiNova Probe PCR Kit

One-Step qRT-PCR (optional): QuantiNova Probe RT-PCR Kit

Assays or panels:

QuantiNova LNA Probe PCR Assays

QuantiNova LNA Probe PCR Reference Assays

QuantiNova LNA Probe PCR Custom Assays

QuantiNova LNA Probe PCR Focus Panels

QuantiNova LNA Probe PCR lncRNA Focus Panels

QuantiNova LNA Probe PCR Custom Panels

QuantiNova LNA Probe PCR Flexible Panels

Applications

QuantiNova LNA Probe PCR Assays and Panels are highly suited for applications including:

mRNA and lncRNA expression analysis, profiling and quantification

Validation of RNA-seq gene expression data

Gene expression profiling

Signal and pathway analysis

Confirming gene expression knockdown by LNA GapmeRs or siRNAs

Biomarker development, including screening, identification and validation of disease-associated biomarkers

Monitoring phenotypic changes related to gene expression