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
Flexible qPCR system
Time-tested QuantiNova chemistry
Easy-to-use data analysis
What you need to get started
|QuantiNova LNA PCR System
||QuantiNova LNA Probe PCR System
|Detection technology||SYBR® Green||FAM-labeled, LNA-enhanced hydrolysis probe
|1||Reverse Transcription||QuantiNova Reverse Transcription Kit||QuantiNova Reverse Transcription Kit|
|2||Mastermix for qPCR||QuantiNova SYBR Green PCR Kit||QuantiNova Probe PCR Kit|
|3||RNA sample quality control (optional)||QuantiNova LNA PCR QC Panels (coming soon)||QuantiNova LNA Probe PCR QC Panels (coming soon)|
|4||Assay or panel||• QuantiNova LNA PCR Assay
• QuantiNova LNA PCR Custom Assay
• QuantiNova LNA PCR Focus Panels
• QuantiNova LNA PCR lncRNA Focus Panels
• QuantiNova LNA PCR Flexible Panels
• QuantiNova LNA PCR Custom Panels
|• QuantiNova LNA Probe PCR Assay
• QuantiNova LNA Probe PCR Custom Assay
• QuantiNova LNA Probe PCR Focus Panels
• QuantiNova LNA Probe PCR lncRNA Focus Panels
• QuantiNova LNA Probe PCR Flexible Panels
• QuantiNova LNA Probe PCR Custom Panels