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Why qNPA™ ?

Quantitative Nuclease Protection Assay (qNPA™)

qNPA Technology offers several important advantages over other gene expression analysis techniques:

No RNA extraction: HTG has successfully run samples from cultured cell lines, primary cells, tissue pieces, FFPE, plants, and bacteria without RNA extraction (For a complete list, click here). Not only does this remove a time-consuming and costly bottleneck in high throughput analysis, but it also reduces a potential source of technical sample variation.

No cDNA synthesis: Traditional methods of gene expression analysis require a reverse transcription of the targeted RNA. qNPA-based analysis does not involve this step, thereby removing another source of potential sample variation.

No RNA amplification or labeling: Coupled with cDNA synthesis, many analysis platforms require a time-consuming RNA amplification and biotin labeling step, adding even more cost and potential sample variation. qNPA uses standard DNA oligonucleotides and S1 nuclease, greatly simplifying the experimental process.

Sensitivity: Single-gene copy sensitivity in as few as 1,000 cells – Measure as few as 1,000 molecules of RNA per well with CV’s of ~10%.

True 47 gene multiplexing: The qNPA ArrayPlate platform will simultaneously measure up to 47 genes in the same sample well. Other multiplexing platforms, such as qPCR, only reliably multiplex 4-6 genes per well, and require extensive development and verification. 

Consistent and reproducible data: Well-to-well CV’s are typically ~10%, far superior to qPCR or microarray data. Data is reliable and consistent, allowing different wells and plates to be compared with confidence and without complex normalization schemes.

Differential expression resolution:qNPA can regularly and accurately detect gene expression changes of as little as 20% (or 1.2 fold) in a plate that has a CV of 10%. qPCR provides reliable resolution of only 50% (or 1.5 fold), which can miss biologically significant changes.