PrimeFlow

Detecting RNA Targets by Flow Cytometry

Invitrogen PrimeFlow RNA Assay expands the capability of flow cytometry to measure RNA. Combine PrimeFlow with immunolabeling of both cell-surface and intracellular proteins using fluorophore-conjugated antibodies. Researchers now can reveal the dynamics of RNA transcription together with protein expression patterns in single cell resolutions

Download brochure

Find PrimeFlow probes for your target 

Principle behind the PrimeFlow RNA assay

  1. Gene specific oligonucleotide target probe set binds to the target RNA sequence 
  2. Preamplifier and multiple amplifier molecules subsequently hybridize to target RNA for signal amplification 
  3. Label probe oligonucleotides conjugated to fluorescent dye detect up to four RNA targets per cell by flow cytometry

Learn more about this technology

Featured PrimeFlow Assay applications

Featured PrimeFlow applications

Evaluate virus RNA in infected cells

Featured PrimeFlow applications

Detect microRNA

Featured PrimeFlow applications

Probe mRNA when no antibody exists to detect the protein

Not finding the application you're looking for? Let us know what you need

Ordering information

Follow the five steps below to begin your PrimeFlow RNA Assay

Instrument Specification
Flow cytometer

Lasers:

  • Blue (488 nm)
  • Yellow-green (561 nm)
  • Red (633 nm)

Filter sets:

  • FITC: 530/20
  • PE-eFluor-610: 610/20
  • APC: 660/20
  • APC-eFluor 780 (APC-Cyanine7): 780/60
Incubator Capable of maintaining temperature at 40 ±1°C
Metal heat block For 1.5 mL microcentrifuge tube, placed inside the validated incubator
Temperature validation kit The Invitrogen ViewRNA Temperature Validation Kit uses a calibrated thermometer to verify the incubator temperature

Optional: Is your target microRNA? This buffer helps ensure you get improved signal and better sensitivity for miRNA.

Optional: Will the assay be processed in a 96-well plate?

  • Four types of probe sets are currently available for RNA detection 
  • Select different types of probe set depending on the RNA expression level 
  • For multiplex analysis with immunolabeling of both cell-surface and intracellular proteins, find fluorophore-conjugated antibodies
Expression level of target RNA Fluorochrome label Laser excitation wavelength (nm) Bandpass filter (nm) Fluorescence channel Probe set type
Low Alexa Fluor 647 633, 635 660/20 APC, Alexa Fluor 647, eFluor 660 Type 1
Low Alexa Fluor 568 561 610/20 PE-eFluor 610, Alexa Fluor 568, PE-Texas Red Type 10
Medium-to-high Alexa Fluor 488 488 530/30 FITC, Alexa Fluor 488 Type 4
Medium-to-high Alexa Fluor 750 633, 635 780/60 APC-Cy7, APC-eFluor 780, Alexa Fluor 750 Type 6

Search for available (in stock) probe sets

Not finding what you're looking for? Go to Custom Branched DNA Probe Set Tool

  • Positive control—use positive control probe sets in every experiment to ensure proper assay performance: RPL13a for human leukocytes and β-actin (ACTB) for mouse tissues; for specific cell types and other recommended genes, refer to Appendix 5 in the product manual of the PrimeFlow RNA Assay Kit
  • Negative control—such as samples with the target specific probe omitted, or samples labeled with a probe against a target not expressed in the cells of interest (e.g., dapB, a bacterial gene); highly recommended

Search probes used for positive and negative controls in probe search tool in Step 4.

Webinars

PrimeFlow resources

Related products

Resources

Flow Cytometry Learning Center—Access flow cytometry educational resources for better experiment planning and execution.

Flow Cytometry Panel Builderbeta—Design your flow cytometry panel with this online tool for a simplified, customizable experience to fit your needs.

Support

Flow Cytometry Support Center—Find technical support recommendations for your flow cytometry workflows, including tips for experimental setup and in-depth troubleshooting help.

Flow Cytometry Protocols

Contact us