Thermo Scientific™ ViewRNA™ HC Screening Assay

Description

The ViewRNA HC Screening Assay is an in situ hybridization method for visualization of up to four target RNAs at a single transcript detection level simultaneously within individual cells. Dual detection with protein is also possible by optimizing experimental conditions. For flexibility, the ViewRNA HC Screening Assay is provided in three modules (each sold separately): ViewRNA HC Screening Assay Kit, ViewRNA HC Screening Signal Amplification Kit, and ViewRNA Probe Sets. All three modules are needed to perform the ViewRNA HC Screening Assay. Note: the ViewRNA HC Screening Assay Kit and ViewRNA HC Screening Signal Amplification Kit are made-to-order kits. Please allow 4-6 weeks for shipment after placing order. 

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