ViewRNA™ HC Screening Assay

Features of ViewRNA tissue assays include:

• Detect up to four RNA targets at single transcription level in cell culture cells in an automated format for 96- or 384-well plates
• Utilize bright Alexa Fluor 488, Alexa Fluor 546, Alexa Fluor 947, and/or Alexa Fluor 750 fluorescent dyes
• Dual detection with proteins possible by optimizing experimental conditions

The ViewRNA assay is a novel RNA in situ hybridization solution based on patent-pending probe set design and proprietary signal amplification technology. ViewRNA assays offer single copy RNA sensitivity in individual cells in a multiplex assay format. Signal amplification is predicated on specific hybridization of adjacent probe set oligonucleotides to a target RNA, resulting in excellent signal-to-noise ratios. Independent but compatible signal amplification systems enable simultaneous detection of up to four target RNAs in a single assay.

The ViewRNA HC Screening Assay is comprised of three modules (each sold separately):

• ViewRNA HC Screening Assay Kit
• ViewRNA HC Screening Signal Amplification Kit
• ViewRNA Probe Sets

How it works

Step 1: Prepare sample. Adherent cells on a solid surface are fixed and permeabilized. Optionally, perform protein detection by following immunocytochemistry protocol.

Step 2: Hybridize probe sets. Gene-specific probe sets hybridize to target mRNAs.

Step 3: Amplify signal. Independent but compatible signal amplification systems enable simultaneous detection of multiple RNAs in a single assay. Distinct sets of PreAmp, Amp, and Label Probe molecules are used to detect different target RNAs.

Step 4: Image. Target mRNAs are visualized using high content imaging systems or standard epifluorescence microscope.

Components of ViewRNA HC Screening Assay Kit

Store following components at 2°C to 8°C:

• Pretease QS: Enzyme in aqueous buffered solution
• Probe Set Diluent QS: Aqueous solution containing formamide and detergent
• Amplifier Diluent QS Aqueous solution containing formamide and detergent
• Label Probe Diluent QS Aqueous solution containing detergent

Store following components at room temperature:

• Protease Stop Buffer Aqueous buffered solution
• Storage Buffer Aqueous buffered solution
• Detergent Solution: Aqueous buffered solution containing detergent
• Wash Buffer Component 1 (Wash Comp 1): Aqueous solution containing detergent
• Wash Buffer Component 2 (Wash Comp 2): Aqueous buffered solution
• Pre-Hybridization Buffer (PreHyb Buffer): Aqueous buffered solution

Components of ViewRNA HC Screening Signal Amplification Kit

Store following components at -20°C:

• PreAmplifier (PreAmp type 4, 6, 8, or 10): DNA in aqueous buffered solution
• Amplifier (Amp type 4, 6, 8, or 10): DNA in aqueous buffered solution
• Label Probe (LP type 4-Alexa Fluor 488, LP type 1-Alexa Fluor 546, LP type 6-Alexa Fluor 647, LP type 10-Alexa Fluor 750): Fluorescent dye-conjugated oligonucleotides in aqueous buffered solution

Notes:

• 1-plex kits only have type 1 reagents
• 2-plex kits have type 1 and 6 reagents
• 3-plex kits have type 1, 4, and 6 reagents
• 4-plex kits have type 1, 4, 6, and 10 reagents