Artist rendition of QuantiGene Plex Assay that uses Luminex xMAP technology for multiplex gene expression analysis

Invitrogen QuantiGene Assays utilize an accurate and precise method for single or multiplexed gene expression quantitation. QuantiGene Assays incorporate branched DNA technology for accurate gene expression profiling. Branched DNA assays allow for the direct measurement of RNA transcripts by using signal amplification rather than target amplification.
 


QuantiGene assay formats

QuantiGene Plex Assay Kits

Multiplex up to 80 RNA targets per single sample using Luminex xMAP bead technology. Design your own panel using our configurator tool or select from one of our preassembled panels.

QuantiGene Singleplex Assay Kits

Accurately measure single RNA transcripts or DNA copy number using a standard luminometer. Use one of thousands of probe sets available or design your own to target custom sequences.


Why QuantiGene?

QuantiGene RNA Assays measure gene expression directly from cell and whole blood lysates or tissue homogenates without the need to purify or amplify RNA. By measuring the RNA at the sample source, the assay avoids biases and variability inherent to extraction techniques and enzymatic manipulations. In addition, this direct measurement helps overcome issues with transcript degradation typically found in samples such as FFPE. Direct measurement is possible because target RNAs are captured through probe hybridization and quantified through branched DNA technology that amplifies the signal. The signal is read using a Luminex instrument for multiplex assays or a luminometer for single targets.

QuantiGene assays demonstrate high data concordance between multiple gene expression platforms (qPCR, microarrays), which allow the ability to switch technologies without having to repeat experiments.

Highlights and key advantages of the QuantiGene assay:

  • Quantitate directly from crude samples—No need for RNA extraction, cDNA synthesis, or PCR amplification
  • Detect low-abundance genes—Exquisite sensitivity allows for the basal measurement of low-expression genes
  • Works with difficult sample types—Works with degraded and cross-linked RNA in FFPE tissues, with fresh or frozen tissues (animal or plant), whole blood, cultured cells, bacteria, and viruses
  • Simple workflow—ELISA-like workflow using 96- or 384-well plates
  • Large inventory of validated genes—Select from over 22,500 genes to create pathways and disease-themed panels.
  • Customization—Customize your panel, and receive your custom panel within 3 weeks.
  • Standardized formats to enable easy scale-up—96-well plate, 384-well plate, and plex-in-plex formats available
     

How the QuantiGene Assays work

Branched DNA technology

QuantiGene Plex and SinglePlex assays utilize branched DNA (bDNA) technology. bDNA technology utilizes sequential nucleic acid hybridization for a unique approach to RNA and DNA quantification by amplifying a reporter signal rather than the template. This measures the transcripts at physiological levels.

First, cells are lysed or tissue samples are homogenized to release the target RNA or DNA. Second, an oligonucleotide probe set is incubated with the target RNA or DNA. During this incubation, the probes cooperatively hybridize to the target. Third, signal amplification is performed via sequential hybridization of the bDNA pre-amplifier, amplifier, and labeled probe molecules to the target. Addition of a chemiluminescent substrate (singleplex assays) or fluorescent reporter (multiplex asssays) generates a signal directly proportional to the amount of target RNA or DNA present in the sample.

A pair of target-specific probe sets (Target Probe), approximately 20 nucleotides in length, hybridizes to contiguous sequences on the target RNA (or DNA). Signal amplification is achieved through successive hybridization of short oligonucleotide sequences to build the bDNA structure (bDNA "tree"), formed by Preamplifiers, Amplifiers, and Labeled Probe, resulting in excellent specificity, low background, and high signal-to-noise ratio.
 

QuantiGene versus qPCR

The QuantiGene assay offers an alternative to qPCR with the ability multiplex up to 80 gene targets directly from cell lysates, whole blood lysates, or tissue homogenates in a convenient plate-based format. While qPCR is considered the gold standard and traditional technology for gene expression analysis to study gene regulation, gene variation, and differential gene expression, a number of steps are required including the isolation of RNA and reverse transcription before performing the qPCR assay. Innovations like Thermo Fisher Scientific’s Cells-to-CT Kits enable measurement of relative gene expression by real-time RT-PCR directly from cultured cells, but direct-from-sample measurement across many different sample types remains a challenge. QuantiGene Assays provide a useful alternative to directly measure RNA expression across multiple different sample types and require minimal method development and optimization.
 

Branched DNA technology

QuantiGene Plex and SinglePlex assays utilize branched DNA (bDNA) technology. bDNA technology utilizes sequential nucleic acid hybridization for a unique approach to RNA and DNA quantification by amplifying a reporter signal rather than the template. This measures the transcripts at physiological levels.

First, cells are lysed or tissue samples are homogenized to release the target RNA or DNA. Second, an oligonucleotide probe set is incubated with the target RNA or DNA. During this incubation, the probes cooperatively hybridize to the target. Third, signal amplification is performed via sequential hybridization of the bDNA pre-amplifier, amplifier, and labeled probe molecules to the target. Addition of a chemiluminescent substrate (singleplex assays) or fluorescent reporter (multiplex asssays) generates a signal directly proportional to the amount of target RNA or DNA present in the sample.

A pair of target-specific probe sets (Target Probe), approximately 20 nucleotides in length, hybridizes to contiguous sequences on the target RNA (or DNA). Signal amplification is achieved through successive hybridization of short oligonucleotide sequences to build the bDNA structure (bDNA "tree"), formed by Preamplifiers, Amplifiers, and Labeled Probe, resulting in excellent specificity, low background, and high signal-to-noise ratio.
 

QuantiGene versus qPCR

The QuantiGene assay offers an alternative to qPCR with the ability multiplex up to 80 gene targets directly from cell lysates, whole blood lysates, or tissue homogenates in a convenient plate-based format. While qPCR is considered the gold standard and traditional technology for gene expression analysis to study gene regulation, gene variation, and differential gene expression, a number of steps are required including the isolation of RNA and reverse transcription before performing the qPCR assay. Innovations like Thermo Fisher Scientific’s Cells-to-CT Kits enable measurement of relative gene expression by real-time RT-PCR directly from cultured cells, but direct-from-sample measurement across many different sample types remains a challenge. QuantiGene Assays provide a useful alternative to directly measure RNA expression across multiple different sample types and require minimal method development and optimization.
 

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