Novel tools for gene expression evaluation
QuantiGene assays are versatile and enable a comprehensive approach to systems biology and translational sciences for validation and quantitation of
biomarkers identified by next generation sequencing, microarray studies or from published literature. QuantiGene assays enable accurate and precise quantitation of RNA, miRNA, and DNA directly from the sample. These direct hybridization assays avoid biases associated with nucleic acid purification, cDNA synthesis and PCR amplification.
- Is qPCR still the gold standard for measuring expression?
- Quantigene Plex Assay: gene expression analysis
- Quantigene Plex Assay: Main steps
- Quantigene Plex Panels
Is qPCR still the gold standard for measuring expression?
qPCR is widely considered to be the gold standard for measuring RNA expression, but with it come several limitations. Researchers in both industrial and academic laboratoires are constantly shifting their assay methods and mindset away from qPCR in favor of higher-content assays that enable them to complete their projects faster.
- … spending too much time purifying RNA?
- … frustrated with the amount of primer design and optimization required?
- … wishing you had lower CVs, better fold-change sensitivity, or more robust Z’-factors?
- … experiencing variations in priming efficiency or reverse transcription?
Imagine what you could accomplish if…
- … RNA purification was NOT necessary, saving you time.
- … reverse transcription was NOT needed, eliminating bias.
- … you could run your custom panel within two weeks of identifying your genes of interest.
- … you could analyze up to 80 targets at once, drastically reducing the number of samples to run.
Quantigene Plex Assay: gene expression analysis
Quantigene® Plex assays provide the fastest and highest throughput solution for quantitative, multiplexed gene expression analysis. Custom panels are designed, quality tested, and shipped within two weeks of your request, and no RNA purification is necessary – just lyse your samples and run the assay.
Quantigene® Plex assays are hybridization-based and use branched DNA (bDNA) signal amplification technology on Luminex xMAP beads. RNA targets are directly quantified with no need for cDNA synthesis or PCR amplification.
Skip RNA purification. Quantigene Plex assays quantitate directly from cell lysates or tissues homogenates, saving you time and minimizing issues caused by contamination or excessive RNA handling.
Reduce 20 qPCR plates to just one Quantigene Plex plate! With true multiplexing capability using Luminex’s xMAP Technology, Quantigene Plex assays measure up to 80 genes of interest in a single well with no cross-reactivity. This capability dramatically increases your efficiency and reduces your workload, while allowing you to normalize against several reference genes all in the same reaction.
Quantitate with confidence. By avoiding the common qPCR pitfalls caused by variations in primer efficiency, reverse transcription, and amplification efficiency, you can trust your Quantigene Plex data. Additionally, the linear readout from Quantigene Plex assays makes data interpretation very simple and meaningful.
Applications. Drug discovery (compound screening, ADME/Tox screening), microarray data validation, biomarker studies using archival tissues and blood, telomere lenght measurement, RNAi gene knockdown efficiency testing, multiplex gene pathway analysis, CNV analysis, iPSC differentiation protocol optimization.
Quantigene Plex Assay: Main Steps
The Quantigene Plex Assays are performed on 96-well plates. The technology is based on DIRECT quantification of RNA targets with branched DNA for multiplexing 3 to 80 RNA targets in a single well.
Step 1. Sample preparation. Samples are lysed to release and stabilize RNA’s. The RNA assay works with a variety of samples such as: cultured cells, human, plant and animal tissues, FFPE tissues, whole blood and PAXGene blood, or purified RNA.
Step 2. Target hybridization. Overnight hybridization in the 96-well plates with the target-specific probe set panels: Capture Extenders [CE], label extenders [LE] y Blocking Probes [BP].
Step 3. Signal Amplification. Signal amplification is achieved using branch DNA (bDNA) technology. A Pre-Amplifier (PreAmp) molecule hybridizes to each pair of Label Extenders, but not to individual probes. Then, multiple Amplifier (Amp) molecules hybridize to each PreAmp. Finally, multiple Label Probe oligonucleotides hybridize to each Amp.
Step 4. Detection. Addition of Streptavidin Phycoerythrin (SAPE) generates a signal that is proportional with the amount of target RNA present in the sample. The signal is read using a Luminex instrument.