The QuantiNova LNA PCR Systems for gene expression analysis merge the precise hot-start mechanism, assay robustness, heightened efficiency and pipetting control of the QuantiNova chemistry with the unmatched specificity of LNA-enhanced oligos. These unique qPCR systems give you the freedom to choose SYBR® Green or an LNA-enhanced hydrolysis probe for RNA and lncRNA detection, and deliver fast and sensitive results with a streamlined protocol based on a universal reverse transcription reaction.
The power of LNA enhancement
LNAs are a class of high-affinity RNA analogs in which the ribose ring is locked in the ideal conformation for Watson-Crick binding, resulting in unprecedented thermal stability upon hybridization to the complementary sequence. Incorporating LNA into oligonucleotides has been shown to improve sensitivity and specificity for many hybridization-based technologies including PCR, microarrays and in situ hybridization. For each incorporated LNA monomer, the melting temperature (Tm) of the duplex increases by 2–8ºC. Incorporating LNA into qPCR primers and probes results in assays with increased binding affinity and enhanced sensitivity.
In addition, LNA oligos can be shorter than traditional DNA or RNA oligos and still retain a high Tm. This is advantageous when designing primers for precise amplification of a specific region of a transcript and also for detecting specific isoforms or SNPs. Plus, intelligent placement of LNA within the primers can increase the Tm between perfect match and mismatch targets, enabling better discrimination of closely related sequences, even those with a single nucleotide difference.
Varying the number of LNAs also allows adjusting the Tm so that all primers have an optimal Tm for the specific qPCR cycling conditions, regardless of the target GC content. Known as Tm normalization, this is especially important for AT-rich transcripts, for which designing DNA primers with sufficient binding affinity is a challenge.
High-performance gene expression analysis
Both the QuantiNova LNA PCR System and the QuantiNova LNA Probe PCR System offer a unique combination of performance and ease-of-use features, but differ in detection strategy.
Both systems use one cDNA reaction for all RNAs – A single universal first-strand cDNA synthesis reaction is used as the template, regardless of the number of targets being profiled. This saves sample, reduces technical variation and pipetting, and saves time.
The QuantiNova LNA PCR System uses two LNA-enhanced, target-specific qPCR primers and SYBR® Green for signal detection.
The QuantiNova LNA Probe PCR System uses an LNA-enhanced, RNA-specific forward primer and a LNA-enhanced, target-specific hydrolysis probe.
LNA-enhanced oligos exhibit high affinity and unrivalled sensitivity and specificity, even in samples with low RNA content.
The advantage of experience
Our expertise in gene expression assay design and LNA oligo enhancement spans more than 20 years and is the foundation for the new QuantiNova LNA PCR Systems. Over 2.6 million predesigned transcript-specific assays for human, mouse and rat genes provide the most sensitive, accurate and effective mRNA and lncRNA analysis. You can also create your own custom assays using our powerful design algorithm, which incorporates over 50 different parameters – all thoroughly lab-validated against highly stringent performance criteria. The advantage to you is assays that work, with no need to spend your time or money on optimization.
Flexible qPCR system
The QuantiNova LNA PCR Systems can be easily tailored to your experimental setup with the choice of individual assays, focus panels for in-depth analysis of 170+ pathways, or fully flexible custom panels. Use the same cDNA reaction to shift seamlessly between individual assays, focus panels or custom panels, depending on your research needs. Predesigned assays are available now as part of our early access program, and the remaining assays and panels will be available soon.
Time-tested QuantiNova chemistry
Both of the QuantiNova LNA PCR Systems were developed to give optimal results and highest reproducibility when used with our trusted QuantiNova chemistry. The hot-start mechanism enables successful reverse transcription, even for difficult templates, such as those with high GC-content or complex secondary structure – ideal for lncRNA analysis. The hot start also allows reaction setup at room temperature, and is robust enough to support automated setup and delayed run of assay plates. Plus, the built-in visual pipetting color indicator helps prevent human pipetting errors. Choose from 2-step or 1-step qRT-PCR reagents and get to your results quickly with our simple-to-follow, 2-hour workflow.
Easy-to-use data analysis
Amplification results collected from experiments using QuantiNova LNA PCR Panels and QuantiNova LNA Probe PCR Panels can be quickly and easily evaluated using our complementary online data analysis tools in the GeneGlobe Data Analysis Center. Simply upload your data, follow the on-screen instructions and get quick insights!
What you need to get started
Both the QuantiNova LNA PCR System and the QuantiNova LNA Probe PCR System require a universal reverse transcription kit (1). Each system requires its own dedicated PCR mastermix (2), combined with your choice of predesigned or custom assays or panels (4). Optionally, RNA quality of samples can be tested with QC panels to ensure safe? use of samples and assays (2).