Search Thermo Fisher Scientific
Search Thermo Fisher Scientific
QuantiGene Singleplex Gene Expression Assays |
QuantiGene Singleplex Gene Expression Assays help provide a streamlined approach to RNA quantification, eliminating the need for reverse transcription or PCR. This assay offers exceptional accuracy and reliability, making it excellent for a wide range of gene expression studies. With its robust performance and simplified workflow, the QuantiGene Singleplex Assay is an important tool for researchers aiming to advance their molecular biology research.
QuantiGene Singleplex assays help provide a fast and high-throughput solution for gene expression quantitation, allowing measurement of a gene of interest in a single well of a 96- or 384-well plate. This hybridization-based assay incorporates branched DNA (bDNA) technology, using signal amplification rather than target amplification for direct measurement of RNA transcripts. With its robust performance and high sensitivity, the QuantiGene Singleplex Assay simplifies workflows with an easy-to-use, ELISA-like process that eliminates the need for RNA purification, reverse transcription, or qPCR (Figure 1). Well suited for a variety of research applications, including biomarker discovery, validation, and molecular diagnostics, this innovative assay enhances research capabilities by offering remarkable accuracy and efficiency in gene expression studies.
QuantiGene assays utilize branched DNA (bDNA) technology, which utilizes sequential nucleic acid hybridization for a distinct approach to RNA and DNA quantification by amplifying a reporter signal rather than the template. First, cells are lysed or tissue samples are homogenized to release the target RNA. Second, the oligonucleotide probe set is incubated with the target RNA and capture plate overnight. During this incubation, the probes cooperatively hybridize to the target and capture probes bound to the plate, capturing the target RNA. Third, signal amplification is performed via sequential hybridization of the bDNA preamplifier, amplifier, and label-probe molecules to the target. Addition of a chemiluminescent substrate generates a luminescence signal directly proportional to the amount of target mRNA present in the sample (Figure 2). The resulting signal is then read on a luminometer, such as the Varioskan ALF Multimode Microplate Reader.
Figure 2. How QuantiGene Singleplex assays work. Samples are prepared to get lysate with target RNA. The target of interest is then hybridized onto the wells of the microplate through a series of steps. Signal amplification and detection is then performed, with the signal read on a luminometer.
View the instructional video showing how to use the QuantiGene Singleplex Assay. Please make sure to carefully read and review your kit’s specific instruction sheet for detailed information before beginning the procedure.
Figure 3. Detection of low-abundance genes. QuantiGene assays were able to detect RNAs from three low-abundance genes in two reference samples: brain RNA and universal human reference RNA. These genes were undetectable using other technologies. This exquisite sensitivity allows for the detection and measurement of low expression genes.
QuantiGene Singleplex assays are exceptional tools for a wide range of applications in molecular biology and research.
Read more about QuantiGene assays and how they are used in the following various immunoassay research areas:
Limit of detection | ≤200 transcripts/assay well |
---|---|
Limit of quantitation | ≤500 transcripts/assay well |
Linear dynamic range | ≥3.5 logs |
Assay CV | ≤10% intra-assay; ≤15% inter-assay |
Accuracy of fold-change | 100% +/- 20% |
Specificity | 99.98% |
Compatible sample types | Cultured cells, bacteria, whole blood, PAXgene blood or dried blood spots, fresh/frozen tissues (animal or plant), FFPE samples, purified RNA |
Assay format | 96-well or 384-well plates |
Targets/well | 1 |
To demonstrate the accuracy of the QuantiGene Singleplex Gene Expression Assay, measurements of twenty transcripts from two reference RNA samples were made and compared to TaqMan data. Reference RNA samples were human brain total RNA and universal human reference RNA [2]. As shown in Figure 5, excellent correlations in fold-change of RNA levels in the reference samples were seen between QuantiGene SinglePlex and TaqMan assays (R2 = 0.965).
In drug discovery, time is money. Fast-track your discovery using the QuantiGene Singleplex High Throughput (HT) Assay, which enables gene expression-based high-throughput screening of thousands of compounds every day. The assay's ELISA-like workflow is easy to automate and does not require RNA purification or extensive assay optimization. The extremely robust QuantiGene Singleplex HT Assay is hybridization-based and uses no enzymes; instead, it relies on branched DNA technology for signal amplification and luminescence for signal readout in a convenient 384-well plate.
Distinct benefits:
High throughput QuantiGene assays have been used by researchers such as Gozgit et al., who investigated the modulation of CYP1B1 levels by PARP7. In the Cancer Cell article, they found that PARP7 is a cancer-specific brake in cytosolic nucleic acid sensing. HARA cells were lysed and then used with the assay kit QS0384 and the corresponding PARP7 probe set [3].
QuantiGene Plex Gene expression assays provide a fast and high-throughput solution for multiplexed gene expression quantitation, enabling the simultaneous measurement of up to 80 genes of interest in a single well of a 96- or 384-well plate.
For Research Use Only. Not for use in diagnostic procedures.