Revvity, 64DSRNAPEG, HTRF dsRNA (IVT) Detection Kit, 500 Assay Points

Overview
In Vitro Transcription or IVT is molecular biology technique used to synthesize RNA sequence from a DNA template. During that process, a DNA template is incubated with RNA polymerase and ribonucleotides, resulting in RNA copies of the template.
While this process aims at producing single-stranded RNA (ssRNA) sequences, it is also prone to generating double-stranded RNA (dsRNA) contaminants. These can arise for different reasons (polymerase pausing, secondary structure formation, template contamination, etc) and are undesirable in the final product.
The end-goal therapeutic applications of IVT are mostly related to gene therapy and mRNA vaccine development or production, and in many cases IVT applications are parts of production processes for patient-intended products. One of the main concerns when running such processes is the QA/QC aspect of it as well as guaranteeing patient safety with the end results. For that reason, dsRNA is a contaminant to monitor with great attention and robust tools, as it is usually a strongly immunogenic molecule that can be picked up by multiple nucleic acid receptors of our innate immunity and trigger violent immune and inflammatory responses in patients. Rapid, convenient, and robust quantification of dsRNA is therefore required at multiple purification steps of the IVT process to ensure maximum elimination of that contaminant.
This HTRF dsRNA (IVT) Kit is intended for the robust detection of dsRNA in such IVT samples. It was developed with IVT applications specifically in mind offers a ready-to-use homogeneous alternative to traditional ELISA and Dot-blot approaches.
How it works
Principle of the HTRF dsRNA (IVT) assay
The dsRNA (IVT) assay measures dsRNA in samples. The assay uses two labeled antibodies: one coupled to a donor fluorophore, the other to an acceptor. In presence of dsRNA in a sample, the addition of these conjugates brings the donor fluorophore into close proximity with the acceptor, and thereby generates a FRET signal. Its intensity is directly proportional to the concentration of the dsRNA present in the sample in a no-wash assay format.

Protocol of the HTRF dsRNA (IVT) assay
The dsRNA (IVT) assay protocol, using a 384-well small volume white plate, is described on the right. 10 µL of sample or standard are dispensed directly into the detection plate for detection by HTRFTM reagents. The antibodies labeled with HTRF fluorophores were added before an overnight incubation. The assay can be run in up to a 1536-well format by simply resizing each addition volume proportionally. Application-Protein Quantification
Brand-HTRF
Detection Modality-HTRF
Product Group-Kit
Sample Volume-10 µL
Shipping Conditions-Shipped in Dry Ice
Target-dsRNA
Target Class-Nucleic Acid
Technology-TR-FRET
Unit Size-500 assay points