QuantiGene™ Plex Assay Kit

Catalog No.	Quantity
QP1013	1 Plate
QP1014	3 Plates
QP1015	10 Plates

Catalog No. QP1013 Supplier Invitrogen Supplier No. QP1013

Description

The QuantiGene Plex Assay Kit contains the generic reagents, buffers, and plates necessary to perform the assay in 96-well plates and must be used with a target-specific QuantiGene Plex panel (sold separately).

The QuantiGene Plex Assay is a gene expression assay that enables multiplexed measurement of up to 80 RNA transcript targets. The assay uses target-specific probes to capture the RNA of interest, and branched DNA technology then amplifies fluorescent signals which are read with a Luminex xMAP instrument.

Features of the QuantiGene Plex Assay include:

• Difficult sample types—works with degraded and cross-linked RNA in FFPE tissues and blood
• True multiplexing—measure up to 80 genes of interest and housekeeping genes in the same well with no cross-reactivity
• Standardized platform—uses 96 or 384-well plate formats compatible with the INTELLIFLEX, Luminex 200, FlexMap3D, and MagPix systems
• Simple workflow—employs an ELISA-like workflow for direct hybridization of transcripts to beads and transcript labeling
• Large inventory of validated genes—select from over 15,000 genes to create pathway and disease-themed panels
• Rapid turnaround for custom panels—if we don't have your gene(s), we can create a custom panel within two weeks

Technology

QuantiGene assays utilize branched DNA (bDNA) technology that has been used for decades in the VERSANT™ diagnostic assays for HIV-1, HCV, and HBV. 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 Luminex xMAP bead set overnight. During this incubation, the probes cooperatively hybridize to the target and capture probes bound to each bead, 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 streptavidin phycoerythrin generates a luminescent signal directly proportional to the amount of target mRNA present in the sample.

Applications

QuantiGene Plex Assay applications include compound screening of drug candidates, biomarker verification, siRNA knockdown efficiency, prospective and/or retrospective analysis of clinical samples, miRNA profiling, microarray validation, predictive toxicology, and detection of translocations and fusion genes.

Specifications

• Format: 96-well
• For Use With (Application): QuantiGene Plex Panel
• For Use With (Equipment): Luminex instrument
• Content And Storage:

  • 36 μL Proteinase K, –20°C
  • 270 μL Blocking Reagent, –20°C
  • 12 mL Label Probe Solution, 2–8°C
  • 12 mL Pre-Amplifier Solution, 2–8°C
  • 12 mL Amplifier Solution, 2–8°C
  • 12 mL SAPE Solution, 2–8°C
  • 12 mL Lysis Mixture, 15–30°C
  • 0.8 mL Wash Buffer Component 1, 15–30°C
  • 13 mL Wash Buffer Component 2, 15–30°C
  • 12 mL10x SAPE Wash Buffer Concentrate, 15–30°C
  • 1 Magnetic Separation Plate, 15–30°C
  • 10 Plate Seals, 15–30°C
  • 1 Hybridization Plate, 15–30°C
  • 2 Pressure Seals, 15–30°C

  Note: Prepared 1x Wash Buffers can be stored for 1 week protected from light at 4°C.

• Product Line: QuantiGene Plex
• Product Type: Multiplex Assay Kit
• Quantity: 1 Plate
• Shipping Condition: Blue Ice
• Sample Type: Purified RNA, cell/blood/FFPE lysates, tissue homogenates
• Detection Method: Fluorescence
• Species: Universal

Frequently Asked Questions (FAQs)

Can samples in TRIzol solution be run in the QuantiGene Plex assay?

No, samples in TRIzol solution cannot be run in the QuantiGene Plex assay. However, RNA purified from these samples can be run in the QuantiGene Plex assay.

Can I use the QuantiGene Plex assay with bacteria?

Although the kits were not originally validated for bacteria, we have guidelines for preparing bacterial samples for QuantiGene Plex assays: http://assets.thermofisher.com/TFS-Assets/BID/Reference-Materials/preparation-bacterial-homogenates-quantigene-plex-assays-protocol.pdf

Do you offer QuantiGene Plex assay custom services where I can send my samples for you to process?

Yes, please reach out to your Technical Sales Specialist for more information or fill out this form: https://www.thermofisher.com/us/en/home/global/forms/life-science/request-quote-quantigene-assays.html

Can I detect genomic DNA using the QuantiGene Plex assay?

Under the hybridization conditions that are used for the QuantiGene Plex assay, genomic DNA remains double-stranded. Therefore, it is not possible for the probes to bind to DNA. The signal from the probe sets comes only from RNA.

Can I use other lysis buffers with QuantiGene Plex assay samples?

We only recommend the use of QuantiGene Lysis Mixture for preparing cell and blood lysates. The Lysis Mixture has been optimized to provide stability of RNA in the lysates. In addition, QuantiGene Lysis Mixture provides the appropriate buffer conditions for the overnight hybridization.

Does Thermo Fisher Scientific currently perform functional testing on the QuantiGene Plex assay probe sets?

For QuantiGene Plex assays, we do perform functional QC of the probe sets using total RNA preparations for the species of interest.

How should I seal the Day 1 plate for Target Probe hybridization for the 384-well QuantiGene Plex assay?

The Day 1 plate can be sealed either manually or using a heat sealer, which is the preferred option. The pressure seal for manual sealing and the heat seal (Cat. No. AB-0757 for Thermo Fisher ALPS 50 V-Manual Heat Sealer, Cat. No. AB-1443A) are provided in the kit. If a different heat sealer will be used, please consult the manufacturer for suitable sealing.

Can I use a heat sealer to seal the plate on Day 2 of the QuantiGene Plex assay?

No, manual sealing using the provided seal is recommended. Heat sealing would deform the plate causing problem in washing.

Should I use control/housekeeping genes in my panel for QuantiGene Plex assays?

Three or more control/housekeeping genes are recommended to normalize the gene expression results with QuantiGene Plex assays. Please refer to the application note "Defining a QuantiGene Plex Assay panel" (https://assets.thermofisher.com/TFS-Assets/LSG/brochures/defining-quantigene-plex-assay-panel-app-note.pdf).

Can I use QuantiGene Plex assays for miRNA or DNA analysis?

miRNA and DNA can only be analyzed with our QuantiGene Singleplex assays, that allow for detection of one target per well.
Here is the link to the QuantiGene Singleplex website: https://www.thermofisher.com/us/en/home/life-science/gene-expression-analysis-genotyping/quantigene-rna-assays/quantigene-singleplex-assay.html

What reagents are included in the QuantiGene Plex assay kit?

The QuantiGene Plex assay kit provides all necessary reagents to perform the assay except Target Probe and Bead mix which are ordered separately. Please refer to the user guide for a list of the reagents included.

What are the proper storage conditions for the QuantiGene Plex assay reagents?

When the QuantiGene Plex assay kit arrives, please follow the instructions in the product sheet to store individual components. The Pre-Amp/Amp/LP Solutions, Bead Mix, SAPE, SAPE Diluent should be stored at 4 degrees C, while the probe set mix, Proteinase K, and blocking reagent should be stored at -20 degrees C. Other reagents should be stored at room temperature.

For QuantiGene Plex assays, what is the recommended temperature for hybridization?

We recommend using Day 1 hybridization temperature of 54 degrees C and Day 2 hybridization temperature of 50 degrees C. Please use QuantiGene Temperature Validation Kit (Cat. No. QS0517) to verify the incubator temperature.

What data analysis software solutions are offered for QuantiGene Plex assays?

The QuantiGene data analysis app is available on the Thermo Fisher Connect Platform for analyzing QuantiGene Plex assay data. For advanced QuantiGene Plex assay data analysis and visualization, the free desktop application software, Applied Biosystems Transcriptome Analysis Console (TAC) software can be used.

What sample types are compatible with QuantiGene Plex assays?

Whole blood, PaxGene blood, fresh tissue or FFPE tissues, cultured cells, and purified RNA have been tested with QuantiGene Plex assays. Other sample types including Tempus blood, bacteria, H&E stained sections, and blood spots are also compatible. Various sample prep protocols and kits are available.

For Research Use Only. Not for use in diagnostic procedures.